Annual Report 2005 - Klinik für Herz- und Gefässchirurgie ...

herzgefaesschirurgie.usz.ch

Annual Report 2005 - Klinik für Herz- und Gefässchirurgie ...

Division of Surgical Research

Annual Report

2005

Department of Surgery

University Hospital Zurich

Switzerland

Division of Surgical Research

Department of Surgery

University Hospital

Rämistrasse 100

CH - 8091 Zurich


2

Photo & Graphic, Division of Surgical Research

Nico Wick and Carol De Simio


Content

Preface 5

1. Organisation 6

2. Research and Development 8

Cardiac Surgery 8

Tissue Engineering 8

Ischemia / Reperfusion Injury 12

Mechanical Circulatory Support 16

Robotic Surgery 18

Visceral & Transplantation Surgery 20

Hepatobiliary & Transplantation Surgery 20

Islet Cell Laboratory 28

Pancreatitis Research Laboratory 30

Colorectal Surgery Laboratory 33

Trauma Surgery 34

Trauma Immunology 34

Osteogenesis Laboratory 37

Innate Immunity Laboratory 40

Computer Assisted Trauma Surgery 43

Musculosceletal tissue conditioning 46

Plastic, Hand & Reconstructive Surgery 48

Tissue Engineering 48

Anti Aging 51

Thoracic Surgery 53

Transplantation Immunology 53

Tissue Engineering 60

Oncology 63

Surgical Intensive Care Units 67

3. Services 70

Surgical skill laboratories 70

Microsurgical laboratory 70

Histology 70

Photo and Graphic services 71

Administration 71

4. Events 72

5. Publications 73

6. Grants 75

7. Awards 78


Prof. Dr. med.

Gregor Zünd,

Head Division of

Surgical Research

Preface

Dear Colleagues

It is my pleasure to present to you the Annual Report 2005 of the Division of

Surgical Research, Department of Surgery at the University Hospital Zurich.

In the year 2005 we have undergone substantial administrative reorganisation

and with the support of all members within the Division organisational problems

could be solved quickly. The new facilities built in 2004 were successfully

implemented in the daily routine and are highly appreciated by technicians and

researchers. The research group of the Division of Surgical Intensive Care

was integrated and could be provided with new laboratories.

The Division’s quarterly internal reports comprising the corresponding

operational expenditures as well as the investments carried out allow for a quality

control by means of timely financial project management. The investments

carried out in the past year included the acquisition of a high performance

liquid chromatography (HPLC) and a reversal microscope.

Both acquisitions are being used by various research groups from our Division

complementing the apparatus pool accordingly.

The weekly lectures held by the Division of Surgical Research at the

University Hospital Zurich are regularly attended by the members of our

Division and other researchers representing an integrative part of the academic

curriculum within the University, the University Hospital and the Federal

Institute of Technology

It is my pleasure to thank all members within our Division and research partners

of the University, University Hospital and Federal Institute of Zurich for the

excellent performance and collaboration last year.

Yours sincerely

Prof. Dr. med. Gregor Zünd

Head Division of Surgical Research


6

Prof. Dr. med.

Michele Genoni

Director Clinic

Cardiac Surgery

Dr. med.

Walter Kuenzi,

Director a.i. Clinic

Plast. - Hand &

Reconstr. Surgery

Prof. Dr. med.

Gregor Zünd,

Head Division of

Surgical Research

Prof. Dr. med.

Pierre - Alain Clavien,

Director Clinic Visceral

& Transpl. Surgery

Prof. Dr. med.

Walter Weder,

Director Clinic

Thoracic Surgery

Prof. Dr. med.

Otmar Trentz,

Director Clinic

Trauma Surgery

Prof. Dr. med.

Reto Stocker,

Head of Intensive

Care Unit

1. Organisation

1.1 Position of the Division of Surgical Research within the

Department of Surgery

Department of

Surgery

Visceral &

Plast. - Hand &

Cardiac Surgery Transplant. Surgery Trauma Surgery Reconstr. Surgery Thoracic Surgery

Division of Surgical Intensive Care Units

Division of Surgical Research


Prof. Dr. med.

Gregor Zünd,

Head Division of

Surgical Research

Susanne Frehner,

Administration

Division of Surgical

Research

Prof. Dr. med.

Simon Philipp

Hoerstrup

Regenerative

Medicine

PD Dr. med.

Guido Wanner,

Robotic Surgery

PD Dr. phil. ll

Rolf Graf,

Co-Head Division of

Surgical Research

Dr. phil ll

Wolfgang Moritz

Ischemia /

Reperfusion

Injuries

PD Dr. phil ll

Rolf Graf

Methodology

1.2 Structural Organisation of the Division of Surgical

Research

Division of Surgical

Research

1.3 Scientific Sections within the Division of Surgical

Research

Division of Surgical

Research

Cardiac Surgery

Research

Visceral & Transpl.

Surgery Research

Trauma Surgery

Research

Plast.-Hand & Reconstr.

Surgery Research

Thoracic Surgery

Research

Surgical Intensive Care

Research

Skill Laboratories

Services

Regenerative

Medicine

Ischemia /

Reperfusion Injuries

Robotic Surgery

Methodology


8

Prof. Dr. med.

Simon Philipp

Hoerstrup

Prof. Dr. med.

Simon Philipp

Hoerstrup

PD Dr. sc. nat.

Stefan

Neuenschwander

Dr. med.

Alberto Weber

cand. med.

Armin Zürcher

Prof. Dr. med.

Gregor Zünd

Dr. sc. nat.

Jens Kelm

Dr. med.

Ian Cummings

Sirpa Price

cand. med.

Silvan Holdener

Prof. Dr. med.

Michele Genoni

Dr. med.

Dörthe Schmidt

Anita Mol

PhD Student

Prof. Dr.

Vijay Kumar

cand. med.

Sandro Imbach

2. Research and Development

2.1 Cardiac Surgery Research

Cardiac Surgery

Research

Ischemia / Reperfusion

Injuries

Circulatory Assist

Robotic Surgery

2.1.1 Cardiovascular Regenerative Medicine (Tissue

Engineering and Cell Transplantation)

Prof. Dr. med. Simon Philipp Hoerstrup

(Director, Regenerative Medicine Program; Professor of Biomedical

Research projects:

■ Human Cell-Based Systems (progenitor, foetal, adult)

■ Extracellular Matrix (proteins, tensegrity)

■ Biomaterials (biodegradable, bio-active)

■ Bioreactor Systems (tension, flow)

■ Biomechanics, Computational Models, Molecular Imaging

■ Animal Models (small and large)

■ Tissue Engineered Cardiovascular Structures (Heart Valves, Vascular Grafts)

■ Microtissue-Based Implants (Myocardium) and Cell Transplantation

■ Molecular Imaging

Cardiovascular

Regenerative Medicine

The Cardiovascular Regenerative Medicine Program comprises Tissue

Engineering and Cell Transplantation and is focused on the development and

in vitro generation of novel, cell based therapies for cardiovascular applications.

These include tissue engineered blood vessels, heart valves as well

as microscale strategies for myocardial regeneration. Presently utilized heart

valve and blood vessel prostheses carry disadvantages for the patients mainly

because non-living, artificial devices are inserted into the human organism.

Tissue engineering enables the in vitro production of autologous, living and

functional replacements with the capacity of growth for congenital application

as an alternative to state of the art artificial replacements. Furthermore,

an additional focus is the development of cell based implants based on the

design of in vitro generated microtissues to improve myocardial functionality

of the diseased heart.


Proof of Heart Valve Tissue Engineering Concept

pre-implantation in-vivo post-explantation

Autologous living tissue engineered heart valve in a sheep model, based on vascular - derived myofibroblasts

and endothelial cells (Hoerstrup et al, Circulation 2000)

Human Heart Valve Tissue Engineering

Human Prenatal Progenitor Cells

Heart valve tissue engineered

from human marrow stromal cells

(Hoerstrup et al. Circulation 2002)

Native EPCs Uptake of ac-LDL vWF staining

Human endothelial progenitor cells isolated from human umbilical cord blood

Schmidt et al. Ann Thorac Surg. 2004

H&E staining CD 31staining vWF staining

Human umbilical cord-derived endothelial progenitor cells seeded onto a living matrix enginereed from

umbilical cord tissue

Schmidt et al. EJCTS 2005


10

Bioreactor Development

Microscale TE for myocardial Regeneration

Achievements 2005

• sized small

• biocompatible materials

• maintenance of sterility

• static conditioning by continuous perfusion

• dynamic conditioning by mimicking diastolic

phase

Mol et al. Ann Biomed Eng 2005

Gravity-mediated production of myocardial microtissue grafts. Myocardial microtissues retain heart-muscle

specifi c morphology (as shown by fl uorescent staining of sarcomeric alpha-actinin), which can be used as

cell grafts or as building blocks to assemble larger tissue patches of any desired shape.

� Fed. Commission for Technology and Innovation Grant (CTI) Hoerstrup SP

� Swiss National Foundation (NFP 46), Hoerstrup SP

� EU Grant Framework Program 6 (Biosys), Horstrup SP

� Bundesministerium für Bildung und Forschung (BMBF Grant), Hoerstrup SP

� Hartmann-Müller-Stiftung, Schmidt D

� Best Oral Presentation 4th Day of Clinical Research 2005, ZKF, Zürich,

Switzerland, Schmidt D


Collaborations

� Department of Biomedical Engineering, Technical University Eindhoven,

The Netherlands

� Center for Integrative Human Physiology, University of Zurich, Switzerland

� Department of Materials, Federal Institute of Technology, Zürich,Switzerland

� Department of Biochemistry, University Zürich, Switzerland

� Department of Mathematics, Federal Institute of Technology, Zürich, Switzerland

� Department of Computational Science, Federal Institute of Technology,

Zürich, Switzerland

� Department of Veterinary Surgery, MSRU Vetclinics, University Zürich, Switzerland

� Department of Cardiology, University Hospital Zürich, Switzerland

� Department of Cardiac Surgery, Children's Hospital, Harvard Medical

School, Boston, MA, USA

� Department of Pathology, Brigham and Women‘s Hospital, Harvard

Medical School, Boston, MA, USA

� Massachusetts Institute of Technology (MIT), Cambridge, MA, USA

� Laboratory for Tissue Engineering, German Heart Centre, Berlin, Germany

� Laboratory for Transplantation Immunology, University Hospital Zürich, Switzerland

� Institute of Chemistry and Applied Biosciences, Federal Institute of Technology

Zürich, Switzerland

� Institute of Anatomie, University of Bern, Switzerland

� Feto-maternal Hematology Research, Department of Obstetrics, University

Hospital Zürich, Switzerland

Selected references:

� Hoerstrup SP, Sodian R, Daebritz S, Wang J, Bacha E A, Martin D P,

Moran A M, Guleserian K J, Sperling J S, Hatsuoka S, Kaushal S, Vacanti

J P, Schoen F J, Mayer J E (2000) Functional living trileaflet heart valves

grown in vitro. Circulation 102(III): 44-49

� Hoerstrup SP, Kadner A, Melnitchouk S, Trojan A, Eid K, Tracy J, Sodian R,

Visjager J, Kolb S, Grunenfelder J, Zund G, Turina M (2002) Tissue

engineering of functional trileaflet heart valves from human marrow

stromal cells. Circulation 106: I-143-150

� Schmidt D, Mol A, Neuschwander S, Breymann C, Gossi M, Zund G,

Turina M, Hoerstrup SP. Living patches engineered from human umbical

cord derived fibroblasts and endothelial progenitor cells. Eur J Cardiothorac

Surg. 2005 May; 27(5):795-800.

� Mol A, Driessen NJ, Rutten MC, Hoerstrup SP, Bouten, CV, Baaijens FP.

Tissue engineering of human heart valve leaflets: a novel bioreactor

for a strain-based conditioning approach. Ann Biomed Eng. 2005

Dec;33(12):1778-88

� Kelm JM, Djonov V, Hoerstrup SP, Guenter CI, Ittner LM, Greve F,

Hierlemann A, Diaz Sanchez-Bustamante C, Perriard JC, Ehler E and

Fussenegger M. Tissue-Transplant Fusion and Vascularization of

Myocardial Micro- and Macrotissues Implanted into Chicken Embryos and

Rats. Tissue Eng. 2006 in press.


12

PD Dr. med.

Reza Tavakoli

Deyan Mihof

PhD

Dr. rer. nat.

Anna Bogdanova

2.1.2 Ischemia / Reperfusion Injury

Antiapoptotic effect of Erythropoietin and Lithium in myocardial

reperfusion injury

PD Dr. med. R. Tavakoli, Dr. rer.nat. A. Bogdanova, Dr. D. Mihof,

Dr. A. Weber, V. Strohmeier

In our previous study the protective effect of antioxidant therapy was investigated

in a heterotopic heart transplant model in the rat. In the present study

we investigate the cardioprotective potential of erythropoietin and lithium salts

against ischemia-reperfusion injury.

Heterotopic heart transplantation is performed from male Lewis donor Rats

(250-300 g) to male Lewis recipient Rats (250-300 g). Recipient rats are divided

in 3 groups: -Group Epo receives 5000U/Kg Eprex (human recombinant

Erythropoietin) 20 min. prior to reperfusion –Group Li receives 10mg/Kg LiCl

20 min.prior to reperfusion –Group C receives no treatment.

In each group recipients are sacrificed at 5 time points: 5, 30, 60, 360 and

1440 min. after reperfusion. Redox status is measured by tissue Gluthation

content (GSH), myocardial injury by Troponin T and water content. Apoptosis

is investigated by measurements of GSK, Akt-cascade, caspase 7.

Twenty four hours after the operation Epo levels in plasma are still exceeding

the basal level 49-fold (3 U/ml vs 61±6 mU/ml). Lithium treatment does not

cause any changes in plasma Epo levels.


Transplanted hearts treated by Epo show no GSH deprivation in contrast to

the grafts of non-treated control animals. Interestingly, Li + is as effi cient as

Epo in preventing reperfusion-induced GSH depletion (Fig 2B). In contrast to

the action at the heart tissue level, Li + treatment causes a decrease in GSH

content of erythrocytes of the recipient animals. Mechanisms of GSH regulation

by lithium are thus tissue-specifi c. Epo treatment is without an effect on

red cell GSH content.

In agreement with our previous fi ndings, swelling of the graft tissue of the control

group 5 min after the onset of reperfusion is due to the Na+ accumulation.

Administration of Epo or Li + abolishes reperfusion-induced swelling and Na +

accumulation.


14

Both Epo and Li+ stimulate GSH production in myocardial tissue and thus

protect the heart from the oxidative stress induced by reperfusion.

Furthermore, stress and tissue damage markers (TnT, ANP, BNP) will be

measured in plasma and fi nally, activity of the members of Akt-cascade will be

monitored and apoptosis intensity in the grafts will be evaluated.


Achievement 2005

� Tavakoli R, Bogdanova A, Ossent P, Grenacher B, Bogdanova N,

Gassmann M, Zund G, Genoni M. Multiparametric characterization of

myocardial ischemia-reperfusion injury. Submitted for publication.

Collaborations:

� Dr L Bestmann, Institute for clinical chemistry, University hospital Zurich

Institute of Veterinary Physiology, Vetsuisse Faculty, University of Zurich

Center of Integrative Human Physiology, University of Zurich

Selected references:

� Wright, G.L. et al. Erythropoietin receptor expression in adult rat cardio-

myocytes is associated with an acute cardioprotective effect for recombinant

erythropoietin during ischemia-reperfusion injury. Faseb J 18, 1031-3 (2004).

� Parsa, C.J. et al. Cardioprotective effects of erythropoietin in the reper-

fused ischemic heart: a potential role for cardiac fibroblasts. J Biol Chem

279, 20655-62 (2004).

� Hausenloy, D.J. & Yellon, D.M. New directions for protecting the heart

against ischaemia-reperfusion injury: targeting the Reperfusion Injury

Salvage Kinase (RISK)-pathway. Cardiovasc Res 61, 448-60 (2004).

� Ananthakrishnan, R., Hallam, K., Li, Q. & Ramasamy, R. JAK-STAT

pathway in cardiac ischemic stress. Vascul Pharmacol 43, 353-6 (2005).

� Chalecka-Franaszek, E. & Chuang, D.M. Lithium activates the serine/

threonine kinase Akt-1 and suppresses glutamate-induced inhibition of

Akt-1 activity in neurons. Proc.Natl.Acad.Sci.U.S.A 20;96, 8745-8750 (1999).

� Jope, R.S. Lithium and GSK-3: one inhibitor, two inhibitory actions, mul-

tiple outcomes. Trends Pharmacol Sci 24, 441-3 (2003).

� Tramontano, A.F. et al. Erythropoietin protects cardiac myocytes from

hypoxia-induced apoptosis through an Akt-dependent pathway.

Biochem Biophys Res Commun 308, 990-4 (2003).


16

PD Dr. med.

Mario Lachat

PD Dr. med.

Markus Wilhelm

Prof. Dr. med.

Rene Pretre

Dr. med.

Hitendu Dave

2.1.3 Mechanical circulatory support

PD Dr. Mario Lachat, Prof. René Prêtre, PD Dr. Markus Wilhelm,

Long-term support

In 2005, the mechanical circulatory support program expanded further. Seven

patients with terminal heart failure, who were in severe low-output syndrome

refractory to medical therapy, were supported with the Berlin Heart INCOR,

a magnetically suspended and intracorporeally implanted axial-flow pump

for left ventricular support, which was introduced in our clinic in November

2004 (fig. 1). The success with this pump was extraordinary. Four patients

were transplanted successfully, three patients are presently still on support

awaiting transplantation. The longest support time is currently 6 months.

Complication rate is very low, and quality of life is excellent. Five patients

could be discharged home, and two patients went back to work while being

on support and waiting for heart transplantation.

The Berlin Heart EXCOR is an extracorporeally located pulsatile pump (fig. 2).

It can be used for biventricular or right ventricular support for which the

INCOR is not the appropriate device. One patient was supported with an

EXCOR right ventricular pump for right ventricular failure following orthotopic

heart transplantation. After five months, the device could be weaned and

explanted, and the patient is currently awaiting hospital discharge.

Figure 1 Berlin Heart INCOR Figure 2 Berlin Heart EXCOR (links: extrakorporale Lage, rechts:

Implantationsprinzip)

Short-term support

In 2005, ten patients were supported with ECMO (extracorporeal membrane

oxygenation) for acute heart and lung failure, respectively. In acute heart

failure, veno-arterial ECMO was implanted in patients with postcardiotomy

heart failure, and as rescue therapy in patients with rapidly developing cardiogenic

shock as bridge to long-term mechanical support or transplantation. In

lung failure, veno-venous ECMO was implanted in patients with ARDS due to

causes such as fulminant pneumonia. ECMO support extended up to 3 weeks

with good mechanical reliability.


Achievements 2005

� Mechanical circulatory support program with excellent bridge-totransplant

rate

Collaborations:

� Levitronics Inc. (Zurich and Boston, USA)

� Berlin Heart (Berlin, Germany)

Selected references:

� Wilhelm MJ, Genoni M, Lachat ML. The Berlin Heart INCOR – A magnet-

ically suspended axial-flow pump for mechanical circulatory support of

patients with end-stage heart failure. Swiss Perfusion 2005;16:19-28

� Salzberg SP, Lachat ML, von Harbou K, Zund G, Turina MI.

Normalization of high pulmonary vascular resistance with LVAD support in

heart transplantation candidates. Eur J Cardiothorac Surg 2005;27:222-5

� Wilhelm MJ, Prêtre R, Noll G, Ruschitzka F, Maggiorini M, Schmid ER,

Genoni M, Lachat ML. A magnetically suspended axial-flow pump (Berlin

Heart INCOR) for mechanical circulatory support: Experience with the

first 5 patients in Switzerland. Kardiovaskuläre Medizin 2005;8 (suppl 8):

S44

� Wilhelm MJ, Lachat ML, Salzberg SP, Prêtre R, Zünd G, Turina MI,

Genoni M. Five year experience with axial-flow pumps as bridge to heart

transplantation: Impact of implant timing on outcome. 4th EACTS/ESTS

Joint Meeting, Barcelona, Spain, 24.-28.09.2005. Book of Abstracts,

p. 239, abstract 135-I.


18

PD Dr. med.

Jürg Grünenfelder

Dr. med.

André Plass

2.1.4 Robotic Surgery and Innovative Technologies

Minimally invasive cardiac surgery

PD Dr. med. Jürg Grünenfelder

� Automated coronary artery bypass operation in the beating heart

� Visualization and computational flow simulation in coronary arteries and

through heart valves

Minimally invasive surgery (MIS) approaches have revolutionized surgery in

the past decade, dramatically reducing morbidity and time required for healing

and rehabilitation. Of the numerous barriers to more widespread implementation

of MIS, probably the most significant is the problem of visualization. This

general need to visualize the surgical site then relates to the needs for modelling

patients, planning procedures, registering these plans to the patient, and

guiding the surgeon in real time. Surmounting these barriers will open the way

to MIS in a wide variety of new procedures.

The focus of this laboratory is the development of precise, adaptable, and

widely deployable computer-integrated systems and devices that make a

wide variety of surgical interventions newly amenable to minimally invasive

cardiac surgical approaches by presenting minimally invasive tools as well

as information about preoperative plans through simulation, anatomy, tool

position and surgical progress to the surgeon in an ergonomic fashion.


Achievements 2005

� Swiss national scientific foundation grant for computer aided and image

guided medical intervention (CO-ME)

Collaborations:

� Departement of Radiology, Universtiy Hospital Zürich (Hatem Alkadhi, MD)

� Laboratory for Thermodynamics in Emerging Technologies, ETH Zürich

(Prof. Dimos Poulikakos)

� Institute of Mechatronic Systems, ZHW (Prof. Charles Brom)

� Physical Electronics Laboratory, ETH Zürich (Prof. Henry Baltes)

Selected references:

� Dynamic cine imaging of the mitral valve with 16-MDCT: a feasibility

study.

Alkadhi H, Bettex D, Wildermuth S, Baumert B, Plass A, Grunenfelder J,

Desbiolles L, Marincek B, Boehm T.

AJR, American J Roentgenol., 2005 Sep;185(3):636-46

� Accuracy of MSCT coronary angiography with 64-slice technology: first

experience.

Leschka S, Alkadhi H, Plass A, Desbiolles L, Grunenfelder J, Marincek B,

Wildermuth S.

Eur Heart J, 2005 Aug;26(15):1482-7.

� Off-pump coronary artery bypass grafting: the Zurich experience.

Tavakoli R, Reuthebuch O, Hofer C, Grunenfelder J, Genoni M.

Heart Surg Forum. 2005;8(4):E246-8.


20

PD Dr. phil ll

Rolf Graf

Clin. Ass. Prof.

Dr. med. Markus

Selzner

Dr. med.

Panco Georgiev

Dr. des.

Ashraf Osman

Dr. med.

Antonio Nocito

Prof. Dr. med.

Alain Clavien

PD Dr. med.

Yinghua Tian

Dr. med.

Harm Hoekstra

Dr. nat.

Jae-Hwi Jang PhD

Udo Ungethüm

2.2 Visceral & Transplant Surgery Research

Visceral & Transpl.

Surgery Research

2.2.1 Hepatobiliary & Transplantation Surgery

Hepatobiliary &

Transplant. Surgery

Islet Cell Laboratory

Pancreatitis Laboratory

Colorectal Surgery

Ischemia / Reperfusion Injury and Liver Transplantation

PD Dr. med. Yinghua Tian; PD Dr. med. Markus Selzner, Dr. med. Nazia Selzner,

Dr. med. Harm Hoekstra, Dr. med. Panco Georgiev, Dr. J.H. Jang,

Dr. des. Ashraf Osman, Dr. med. Antonio Nocito

Ischemia/reperfusion injury of the liver represents an important problem in

major hepatic surgery, liver transplantation, shock and trauma. Two types of

injury have to be differentiated: (1) warm ischemic injury, occurring during liver

resection, trauma and shock and (2) cold ischemic injury, occurring during

organ preservation. Our group has a longstanding interest in the cellular

processes leading to these two types of injuries. Additional factors affecting

the outcome after ischemia and reperfusion are the presence of steatosis,

cholestasis, cirrhosis and the age of the liver.

To assess ischemic injury in the fatty liver, intravital microscopy was used. We

could show that the type of steatosis (macro- versus microsteatosis) has a

strong impact on various parameters of liver perfusion and activity of leukocytes.

Protective strategies e.g. ischemic preconditioning and intermittent clamping

could improve these parameters. These results are in accordance with a

recent publication in which the impact of steatosis on liver injury after ischemia

and reperfusion was demonstrated (Selzner et al 2006).

Currently, investigations into the ability of the old liver to tolerate ischemic

injury are performed. Results indicate that old livers are more prone to tissue

injury which can be partially corrected by supplying glucose prior to a surgical

insult. Here we could show that the ATP content of the liver was dramatically

increased after a supplement of glucose. Concurrently, the injury was strongly

reduced.

Against the background of platelets mediating sinusoidal endothelial cell

apoptosis in cold hepatic ischemia, we investigated in a further project the role

of platelets in normothermic ischemia and reperfusion injury.

We therefore used a model of platelet dysfunction and immune thrombocytopenia.

Platelet aggregation in mice was inhibited by Clopidogrel feeding.

Immune thrombocytopenia was induced by intraperitoneal injection of anti-

CD41 antibody. Subsequently, all mice were subjected to sixty minutes of

partial hepatic ischemia and various time points of reperfusion.


Hepatic injury was determined by measurement of transaminase levels and by

histological analysis of necrotic area and leukocyte infi ltration. In addition liver

repair after ischemic injury was investigated in platelet depleted animals and in

mice lacking peripheral serotonin, since we have recently shown that platelet

derived serotonin mediates liver regeneration.

In this study we could show that neither inhibition of platelet aggregation nor platelet

depletion led to an improvement of I/R injury. However, liver regeneration and

remodelling were signifi cantly impaired in platelet depleted animals. Conversely,

mice lacking peripheral serotonin were only defi cient in liver regeneration.

From these results we concluded that platelets have no direct impact on the

pathogenesis of normothermic I/R-injury but mediate tissue remodelling and

liver regeneration. In addition, we demonstrate that platelet derived serotonin

is also a specifi c mediator of regeneration in the postischemic liver.

Intra-hepatic ATP contents in young (• ) and old (▪ )

mice subjected to ischemic injury alone, in old mice

with D-glucose injection prior to ischemia (�), and

old mice receiving D-glucose injection plus ischemic

preconditioning prior to ischemic injury (▲).

Administration of D-glucose resulted in a significant

increase in intrahepatic ATP levels after ischemia/

reperfusion in old mice. Combining preconditioning

and glucose administration resulted in an even more

dramatic increase in intrahepatic ATP contents.

The mouse orthotopic liver transplantation (OLT) model was further developed:

it was shown that 30% transplantation of a mouse liver results in a small-for-size

syndrome with high morbidity and mortality. Various approaches to protect the

small-for-size liver syndrome resulted in the detection of a mechanism involving

TNF-α and Kupffer cells. Indeed, application of pentoxifylline, a drug routinely used

in clinical therapy for thrombosis, was successful in reverting the small-for-size

syndrome. Pentoxifylline suppresses TNF-α synthesis and has rheological activities,

improving blood circulation (Tian et al 2006 PNAS, in press).

Histological analysis of graft tissue after transplantation.

H&E staining of partial OLT grafts was performed 2 days after transplantation. In contrast to untreated 30%

partial OLT grafts, which revealed diffuse microvesicular steatosis, only mild macrovesicular steatosis was

found in 30% partial OLT grafts after treatment with pentoxifylline and GdCl 3 (impairs Kupffer cell function) and

in TNFR-1(-/-)grafts (disrupts the TNFα signalling pathway).

The effect of toxic bile on cold reperfusion injury was assessed. In a model of

phospholipid depletion (Mdr2 knockout mice) we could show that altered bile

composition affects the pathology of the bile duct and concurrently leads to increased

liver injury after transplantation (Hoekstra et al Hepatology 2006, in press).


22

Many patients are affected by cholestasis prior to surgery. Hence the question

was raised whether this condition has an impact on surgical interventions. A

model of total bile duct obstruction was established. Mice used in these experiments

exhibited strongly increased levels of bilirubin, indicators of cholestasis.

We could show that cholestatic mice were protected from ischemic injury. The

mediator of the protective effect is systemic which could be experimentally

shown by selective obstruction of individual liver lobes.

In further experiments, the development of cholestasis in a complete bile-duct

ligation model was established. Markers of injury and bile-duct proliferation

were established and quantifi ed. This long-term experiment will provide the

basis for subsequent studies into the mechanisms of injury during cholestasis.

An additional project is currently focusing on the cirrhotic liver. We are trying

to understand whether surgical and pharmacological strategies can protect

the cirrhotic liver from ischemic injury. A model of liver cirrhosis was used that

exhibits extensive tissue rearrangement (CCl4 ).

Achievements 2005

Scientific

� We used a unique model of arterialized partial liver transplantation in

mice. We could show that recipients of small-for-size livers (30%) have

an improved survival rate after treatment with pentoxifylline.

� Establishment of mouse models of cholestasis

� The role of energy charge in aging livers: pre-treatment of animals with

glucose reduces ischemic injury of the liver.

� Investigations into the mechanism of ischemia reperfusion injury in fatty

livers led to the conclusion that these livers exhibited reduced sinusoidal

perfusion and impaired leukocyte function.

� Grant awarded by the SNF.

Personnel

PD Dr. Tian received a poster prize from the ZKF (4 th Day of Clinical Research)

Collaborations:

� PD Dr. W. Jochum, Institut für Klinische Pathologie, UniversitätsSpital

Zürich

� PD Dr. B. Ludwig , Forschungsabteilung, Kantonsspital St.Gallen

Selected references:

� N Selzner, HA Rüdiger, R Graf, PA Clavien. Protective strategies against

ischemic injury of the liver. Gastroenterology 2003;125:917-936.

� Dahm F, Georgiev P, Clavien PA. Small-for-Size Syndrome After Partial Liver

Transplantation: Definition, Mechanisms of Disease and Clinical Implications.

Am J Transpl 2005;5(11):2605-2610.

� N. Selzner, M. Selzner, W. Jochum, R. Graf, B. Ammann-Vesti, P.-A. Clavien.

Mouse Livers with Macrosteatosis are more Susceptible to Normothermic

Ischemic Injury than those with Microsteatosis. J. Hepatology 2005, in press.

� Y. Tian, W. Jochum, P. Georgiev, W. Moritz, Rolf Graf, and P.A. Clavien.

Kupffer Cell dependent TNF-α Signaling mediates injury in the Arterialized

Small-for-Size Liver Transplantation in the Mouse. PNAS 2006, in press


Dr. med.

Nazia Malekkiani

Selzner

Dr. med.

Stefan Heinrich

Dr. med.

Mickael Lesurtel

Dr. med.

Katarzyna Katarzyna Furrer

Liver regeneration

Dr. med. Nazia Selzner, Dr. med. Mickael Lesurtel, Dr. med. Stefan Heinrich

Dr. med. K. Furrer

The liver is the only solid organ with the capacity to regenerate its volume after

major tissue loss. Previous studies have shown that platelets are involved in a

critical step during regeneration. To identify mediators of hepatocyte proliferation,

soluble factors generated by platelets were investigated. One factor with

the capacity to induce proliferation, serotonin, was analyzed. Animal studies

with serotonin agonists were used to prove that serotonin may be a key mediator.

Thrombocytopenic animals which do not regenerate their liver after 70%

liver resection, received the serotonin agonist. This could reconstitute hepatocyte

proliferation. Further studies, including knock-out mice for an enzyme

involved in serotonin biosynthesis, could show that serotonin was a decisive

factor in the induction of hepatocyte proliferation.

Demonstration of reduced liver regeneration in partially hepatectomized mice after depletion of platelets i.e.

after immune thrombocytopenia (A).

Hepatocyte proliferation could be recovered by injection of a serotoinin agonist (DOI).

B: Relative regulation of transcripts coding for serotonin receptor subtypes after partial hepatectomy.

C: Mice treated with serotonin antagonists prior to and after resection. The 5HT2A receptor antagonist completely

blocked regeneration, supporting the conclusion that serotonin is critically involved in liver regeneration.


24

Achievements 2005

Scientific

� Demonstration that serotonin is a key mediator in liver regeneration

Collaborations:

� PD Dr. Odermatt, Labor für Molekulare Diagnostik, Institut für Klinische

Pathologie

� PD Dr. W. Jochum, Institut für Klinische Pathologie

� Prof. M. Bader, Max Delbrück Center for Molecular Medicine, Berlin

Selected references:

� Selzner N, Selzner M, Odermatt B, Tian Y, Van Rooijen N, Clavien PA.

ICAM-1 triggers liver regeneration through leukocyte recruitment and

Kupffer cell-dependent release of TNF-alpha/IL-6 in mice.

Gastroenterology. 2003 Mar;124:692-700.


PD Dr. med.

Philipp Dutkowski

Dr. med.

Katarzyna Furrer

Hypothermic oxygenated rat liver perfusion

PD Dr. Philipp Dutkowski, Dr. med. Katarzyna Furrer

During cold storage of livers cellular components deteriorate resulting in energy

depletion and loss of function. In addition, the mitochondrial Redox state is

affected that greatly reduces the ability of the mitochondrium to transfer energy.

Tissue damage from cold storage is further aggravated by rewarming and

reperfusion. The combined tissue injury eventually leads to a loss of the graft.

Extending or rectifying some of the cellular components might allow to prolong

cold ischemia time and may improve the viability of the grafts. In experiments

using isolated perfused rat livers, the deterioration of cold stored livers could

be demonstrated. A method was developed that could partially rectify this

damage: hypothermic oxygenated perfusion (HOPE) is an approach by which

the livers can be recharged during the last phase of cold storage. The liver

is perfused with an oxygenated, modified UW solution. We could show that

machine perfusion improved several parameters including lipid peroxidation,

bile flow, energy charge, total glutathione and LDH release. Structurally, these

livers exhibited less tissue damage as assessed by electron microscopy.

Furthermore, apoptosis was reduced suggesting that central mechanisms of

tissue destruction were down regulated.

Isolated rat liver perfusion system. The central part

consists of temperature controlled chamber to host

the liver. Circulating perfusion fl uid is pumped through a

reservoir system to keep constant pressure. Sampling

of the perfusate and bile production can be

accessed any time.

Achievements 2005

Scientific

� Demonstration that HOPE can prevent tissue injury in the explanted

liver

Personnel

� PD Dr. P. Dutkowski won an award from the SGC in Zürich

Selected references:

� P. Dutkowski, R. Graf, PA Clavien

Rescue of the cold preserved liver by hypothermic oxygenated machine

perfusion. Am. J. Transplantation 2006 (in press).


26

Dr. med.

Stefan Heinrich

Dr. med.

Daniel Dindo

Dr. med.

Felix Dahm

Dr. med.

Michelle de Oliveira

Oncology

Dr. med. Felix Dahm, Dr. med. Stefan Heinrich, Dr. med. Daniel Dindo,

Dr. med. Michelle de Oliveira

Liver metastases of colorectal cancer are frequent indications for liver resections

in clinical practice. Liver resection is followed by a regenerative response of

the remaining liver. This process of liver regeneration involves growth factors

and cytokines which are also implicated in the promotion of tumour growth invitro.

Therefore the interplay of liver regeneration and colorectal liver and lung

metastases was investigated in a mouse model. Animals were injected with a

syngeneic colorectal cancer cell line, followed by liver resection (PH) or portal

vein ligation (PVL) 7 days later. Liver regeneration and tumour burden were

evaluated 7 days after surgical interventions. We found hepatic tumour load to

be significantly higher after PVL than after PH, while the growth of lung metastases

was not affected by either procedure. We could also show that the presence

of liver metastases reduces liver regeneration after resection, while the

regenerative response was normal after portal vein ligation. Furthermore we

could show that the cytokine TGFβ is produced by tumour cells in-vitro and in

tumor tissue in-vivo. This observation might be the explanation for decreased

liver regeneration in metastatic disease. These observations have triggered

several new experiments investigating the underlying pathomechanisms

(Heinrich et al, J.Hepatology 2006, in press)

Another project focuses on the role of sphingolipids in cancer cell death and

survival. Sphingolipids are a complex class of lipids that not only constitute

cellular membranes, but also play an important role in intra- and extracellular

signalling. The sphingolipid ceramide plays an important role in the induction

of apoptosis. By using liquid chromatography and mass spectrometry we

could show that specifically targeted cationic ceramide analogues accumulate

in mitochondria in vitro. These compounds exert cytotoxic effects on a

variety of cancer cell lines. We studied the human colorectal cancer cell line

SW403 in detail and showed a decreased mitochondrial membrane potential,

cytochrome c release and cytoplasmic caspase activation. Cells reacted to

the ceramide compound by an overall decrease of endogenous ceramide,

and a rapid rise of sphingosine-1-phosphate, a sphingolipid counterplayer

of ceramide. In vivo we could establish a tolerable dosing regime along with

pharmacokinetic behaviour of the ceramide compound. Most importantly,

ceramide treatment reduced the growth of subcutaneous tumour metastases

in vivo. Targeting mitochondria with ceramide analogues offers a promising

new approach to cancer treatment.

In another project we have begun to analyse the effect of chemotherapy on liver

morphology and especially on the liver’s ability to regenerate after hepatectomy.

This study was prompted by the fact that patients with liver metastases receive

chemotherapy followed by surgical resection of residual tumours. There is

anecdotal evidence to suggest regenerative defects of the liver after chemotherapy.


Treatment of s.c. tumours

Assessment of tumour growth of mice treated with either Doxicycline, with a long chain ceramide analogue

(LCL-30) or with a combination of both.

Achievements 2005

Scientific

� Colorectal cancer cells reduce liver regeneration by producing TGFβ

� Dose-finding, pharmacokinetics and proof of efficacy of a long-chain

ceramide analogue in vivo

Personnel

� Dr. D. Dindo received an award for his lecture at the annual meeting of

the Swiss Society of Surgery.

� Dr. F. Dahm received a grant from the Sassella Foundation.

� Dr. S. Heinrich received the Research Prize of the Swiss Surgical

Society and a grant from the Cancer League of Zurich.

Collaborations:

� Prof. Dr. Y.A. Hannun and Prof. Dr. A. Bielawska, Department of Biochemistry

and Molecular Biology, University of South Carolina,

Charleston, USA

� PD Dr. W. Jochum, Department of Pathology, University Hospital Zurich

Selected references:

� Selzner M, Bielawska A, Morse MA, Rüdiger HA, Sindram D, Hannun YA,

Clavien PA. Induction of apoptotic cell death and prevention of tumor

growth by ceramide analogues in metastatic human colon cancer. Cancer

Research (2001)

� Selzner N, Selzner M, Graf R, Ungethuem U, Fitz JG, Clavien PA. Water

induces autocrine stimulation of tumor cell killing through ATP release

and P2 receptor binding. Cell Death and Differentiation (2004)


28

PD Dr. med.

Markus Weber

Lu Tuyet Trinh

Dr. phil. ll

Wolfgang Moritz

2.2.2 Islet Cell Laboratory

The fate of grafted islets in the immediate posttransplantation period

PD Dr. med. Markus Weber, Dr. Wolfgang Moritz

Our main research interest is focused on islet cell transplantation in particular

to develop strategies to improve its efficacy and is performed in close collaboration

with the Endocrinolology and Diabeteolgy Unit of the University Hospital.

For the last five years, islet transplantation has become a widely used therapy

for patients with type 1 diabetes mellitus. Islets are isolated from pancreata of

heart-beating donors and transplanted into the liver of diabetic recipients by

transhepatic portal catherization. Unfortunately, the requirement of donor tissue

is quite high (2-3 pancreata) in order for a diabetic patient to become insulin

independent. The reasons for such a high demand are manifold; insufficient

isolation efficiency and graft rejection are certainly to mention. It has been suggested,

that due to their devascularized state, transplanted islet would potentially

suffer from near-ischemic conditions and consequently undergo massive

cell death. We therefore are investigating strategies to interfere successfully

with the deleterious effects of hypoxia and nutrient deprivation for the immediate

posttransplantation period until complete revascularization of the transplant

has taken place.

Rat islet after syngeneic, intrahepatic transplantation: (A) Selective loss

of glucagon expressing alpha-cells (red). (B) Microvasculature (red) predominantly

formed at the islet’s periphery. Insulin staining (green) of the

islet graft is markedly decreased in diabetic rats (D/Tx) when compared to

normoglycemic recipients (N/Tx).


Achievements 2005

� In order to evaluate the fate of intrahepatically transplanted islets, we performed

a series of syngeneic islet transplantation in diabetic rats.

By the choice of a subtherapeutic islet mass we aimed to simulate transplantation

of marginal donor tissue. As opposed to in vitro studies under

hypoxic conditions, histological examination of islet grafts did not reveal

obvious signs of massive tissue loss, nor signs of cell death by necrosis

or apoptosis. This was also confirmed by the assessment of total insulin

positive beta-cell area in histological sections, which did not change in

the course of the first 14 days after transplantation. However, we observed

a selective loss of glucagon producing alpha-cells, while graft revascularization

was predominantly confined to the islet periphery and accompanied

by strong stroma formation. Cellular insulin content seemed to be decreased

when compared to native pancreatic islets or islets transplanted

into normoglycemic recipient rats. Post transplantation insulin treatment

improved graft performance which was paralleled by restored insulin

stores. Despite their initial avascular state, intrahepatically transplanted

islets do not undergo substantial cell death but rather loose their secretory

capacity due to exhaustion. Hence, islet graft function is negatively

affected by the diabetic state and therefore largely dependent on the

maintenance of normoglycemia, particularly in the immediate early phase

after transplantation. Our observations underline the need of a tight

blood glucose control post-transplantation, especially when only marginal,

subtherapeutic graft tissue was available for transplantation.

This project is supported by the Swiss National Science Foundation.

Additional funding has been received by the Olga-Mayenfisch,

Hartmann-Müller and the Hermann-Klaus Stiftung.

Collaborations:

� The research project is also part of a close collaboration with PD Dr.

Lehmann and Dr. Züllig of the Unit of Endocrinology and Diabetology in

Internal Medicine of the University Hospital Zürich and Prof. Max Gassmann

of Veterinary Physiology of University of Zürich.

Selected references:

� Moritz W., Meier F., Stroka D. M., Giuliani M., Kugelmeier P., Nett P.C.,

Lehmann R., Candinas D., Gassmann M., Weber M. (2002) Apoptosis in

hypoxic human islets correlates with HIF-1 alpha expression. FASEB J.

2002 May; 16:745-7

� Giuliani M, Moritz W, Bodmer E, Dindo D, Kugelmeier P, Lehmann R,

Gassmann M, Groscurth P, Weber M. Central necrosis in isolated hypoxic

human pancreatic islets: Evidence for post-isolation ischemia.

Cell Transplant: 14: 67-76, 2005


30

PD Dr. phil. II

Rolf Graf

Dr. med.

Li K. Sun

Dipl. phil. II

Theresia

Reding Graf

cand.med.

Federico

Storni

PD Dr. med.

Daniel Bimmler

Dr. rer. nat.

Franco Fortunato

Martha Bain

2.2.3 Pancreatitis Research Laboratory

PD Dr. Rolf Graf, PD Dr. med. Daniel Bimmler

The role of COX-2 in chronic pancreatitis

Dr. med. Li-Kang Sun, Martha Bain, Dipl. phil. II Theresia Reding Graf,

cand. med. Federico Storni

Our laboratory has a long standing interest in the pathophysiology of chronic

pancreatitis.

We used a spontaneous model of pancreatic inflammation and fibrosis (WBN/

Kob rat) to test whether a COX-2 inhibitor reduces inflammation and fibrosis.

During the early phase of pancreatitis, inflammatory cells increase dramatically

in untreated rats while in COX-2 inhibitor fed rats they were reduced in

number. One of the predominant cell types, the macrophage, was significantly

less abundant in treated rats and appears to affect the course of pancreatitis.

Concomitant activation of pancreatic stellate cells was strongly inhibited suggesting

that pancreatic fibrosis is targeted by the COX-2 inhibitor. Parameters

of acute and chronic inflammation were significantly down regulated e.g.

TNF-α, Il-6 MCP-1 and TGF-β.

These observations led us to test whether COX-2 might be involved in chemotaxis.

Therefore, we performed cell culture experiments to demonstrate

that prostaglandins (down-stream products of COX-2) modify the behaviour

of macrophages. In chemotaxis chambers we show that the directional

response of a mouse macrophage cell towards a chemotactic signal was

strongly inhibited in the presence of a COX-2 inhibitor.

The relationship of COX activity (PGE 2 ) with macrophage recruitment in

pancreatitis was further investigated. MCP-1, a chemoattractant, was found

predominantly in pancreatic acinar cells in the rat model. To simulate interactions

of macrophages and acinar cells, we used a mouse macrophage cell

line (RAW 264.7) and a rat acinar cell line (AR42J). We then verified that

PGE 2 receptors EP 1-4 are actually expressed in AR42J cells. The expression

of MCP-1 was regulated by TNFα and PGE 2 in pancreatic cells. In the presence

of PGE 2 , TNFα dependent expression of MCP-1 was significantly higher than

with TNFα alone. TNFα regulated the expression of itself in this cell line. In

the presence of PGE 2 , expression levels of TNFα were further enhanced.

Activated macrophages induced the secretion of pancreatitis-associated protein

in the AR42J cell. The presence of acinar derived molecules caused a

markedly increased directional migratory response in macrophages.

We conclude that the chemical interactions of macrophages and acinar cells

might be a key step in establishing the chronic phase of pancreatitis.


Acinar cell damage (*) leads to the nearby accumulation of activated macrophages (#) due to chemoattraction,

e.g. exerted by MCP-1 which is secreted in the vicinity of injured acinar cells.

Activated macrophages produce and shed various cytokines, chemokines, prostaglandines and TGF-β. The

latter stimulates the activation of stellate cells which consequently synthesize and deposit collagen fibers,

eventually leading to fibrosis.

COX-2 inhibitors interfere with different processes: they inhibit chemoattraction and the secretory activity of

macrophages, especially the secretion of TGF-β.

In our WBN/Kob rats we have shown that infiltration of macrophages and subsequent fibrosis are significantly

reduced and delayed if rofecoxib, a COX-2 inhibitor, is administered.

Inset: In vitro experiments using a macrophage cell line (RAW 264.7) demonstrated a similar effect of rofecoxib:

the directed migration of macrophages caused by the chemoattractant fMLP was significantly reduced

with rofecoxib.

We therefore hypothesize that the infiltration of activated macrophages represents an essential step in the

pathogenesis of chronic pancreatitis.

Full arrow: substances secreted by a cell. Broken arrows: positive regulation. Line with a dash at the end:

negative regulation.

Acute pancreatitis in the alcoholic pancreas

Dr. Franco Fortunato

Alcohol is an important factor in the etiology of pancreatitis. To assess whether

alcohol modifies the response to noxious stimuli, we analyzed the pancreata

derived from animals after chronic alcohol feeding and a single dose of LPS

(lipopolysaccharide). There was a dose dependent response of tissue injury

in animals injected with LPS. Concurrent alcohol feeding aggravated the

response. We could show that apoptosis was significantly reduced in alcohol

fed rats. In the presence of LPS, this was further inhibited. We conclude that

inhibition of apoptosis after alcohol exposure may lead to a switch to necrotic

cell death. This in turn might activate a stronger inflammatory reaction, which

we could demonstrate by quantification of macrophages (Fortunato et al, Am.

J. Physiol 2006, in press).


32

Achievements 2005

Scientific

� We have been able to show that chemical signals from macrophages

and acinar cells promote chronic activation and may represent a mechanism

of chronic inflammation and fibrosis.

� COX-2 is involved in macrophage recruitment.

� Apotosis is inhibited by alcohol and tissue injury is switched to a

necrotic type of cell death. Publication in Am. J. Physiology.

Collaborations:

� Dr. Aurel Perren, Institut für klinische Pathologie, Universitätsspital Zürich

� Dr. Pia März & Prof. U. Otten, Physiologisches Institut, Universität Basel

� PD Dr. Marius Keel & Dr. Luc Härter, Klinik für Unfallchirurgie,

Universitätsspital Zürich

� Dr. Martin Hersberger, Institut für Klinische Chemie, Universitätsspital

Zürich

� Dr. George Scheele, Institute of Genomic Medicine, LaJolla, Ca. USA

� Dr. Robert DeLisle, Anatomy and Cell Biology, University of Kansas

School of Medicine, Kansas City, KS

Selected references:

� Graf R, Schiesser M, Reding T, Appenzeller P, Sun LK, Fortunato F,

Perren A, Bimmler D. Exocrine Meets Endocrine: Pancreatic Stone

Protein and Regenerating Protein-Two Sides of the Same Coin.

J Surg Res 2005 (epub ahead of print)

� Reding TV, Bimmler DR, Perren A, Sun LK, Fortunato F, Storni F, Graf

R. A selective COX-2 inhibitor suppresses chronic pancreatitis in an animal

model (WBN/Kob rats): significant reduction of macrophage infiltration

and fibrosis. Gut 2005 (epub ahead of print)

� Fortunato F, Deng X, Gates LK, McClain CJ, Bimmler D, Graf R, et al.

Pancreatic Response to Endotoxin after Chronic Alcohol Exposure:

Switch from Apoptosis to Necrosis? Am J Physiol Gastrointest Liver

Physiol 2005. (epub ahead of print)


Dr. med.

Franc Hetzer

Dr. med. L. Sun

Clin. Ass. Prof.

Dr. med. Dieter

Hahnloser

2.2.4 Colorectal Surgery

Dr. Franc Hetzer MD, Dr. Dieter Hahnloser MD, Dr. L. Sun, MD,

PD Dr. Nicolas Demartines

Haemorrhoids are a frequent and disturbing benign disease of the anus. The

pathogenesis of haemorrhoids is multifactorial including a stasis in the

haemorrhoidal venous plexus causing inflammation and dilatation of the

venous membrane and resulting in chronic bleeding, itching and pain. The

inflammation is triggered by intracellular- (ICAM) and vascular cell adhesion

molecules (CVAM) followed by liberation of prostaglandines. Flavinoid

(Daflon © ) inhibits these adhesion molecules and therefore reduces prostaglandine

levels. In vitro and animal studies have demonstrated increased

vasoconstriction of the venous plexus, reduced capillary leakage and reduced

prostaglandine levels E 2 , F 2α and thromboxan B2 after Daflon © . Several

clinical studies have shown that Daflon © significantly reduces the classical

symptoms of acute and chronic bleeding.

We are investigating in a prospective randomised trial of patients requiring

hemorrhoidectomy the influence of Daflon © perioperatively vs. placebo on the

prostaglandine concentration within symptomatic haemorhhoids. We hope to

demonstrate the local effect of orally taken Daflon © on prostaglandine synthesis

in hemorrhoidal tissue and therefore to elucidate the role of prostaglandines

in the pathogenesis of this frequent disease.


34

PD Dr. med.

Marius Keel

PD Dr. med.

Marius Keel

Dr. med.

Ludwig Labler

Ursula

Steckholzer

Prof. Dr. med.

Otmar Trentz

Dr. rer. nat.

Luc Härter

Dr. med.

Ladislav Mica

2.3 Trauma Surgery Research

Trauma Surgery

Research

2.3.1 Trauma Immunology

Trauma Immunology

Osteogenesis

Innate Immunity

Computer Assisted

Trauma Surgery

Musculosceletal tissue

conditioning

Neutrophil activation and accelerated angiogenesis in vacuum assisted

closure-treated wounds after trauma

PD Dr. med. Marius Keel MD, Dr. rer. nat. Luc Härter

Wound healing is a complex chronologically and spatially organized process

initialized by a local inflammation and infiltration of neutrophil granulocytes

(PMN) and macrophages. These infiltrating leukocytes set the stage for later

processes which finally lead to remodeling of the vasculature and the affected

tissue. The management of large, possibly infected wounds poses a major

challenge in trauma surgery. In the past many different methods of temporary

wound closure have been exploited. A current standard is the wound cover by

Epigard ® and the Vacuum Assisted Closure (V.A.C. ® ) system. The V.A.C. ® -

sytem is a temporary wound cover with a polyvinyl foam sealed with a polyurethane

foil. In the clinical practice the V.A.C. ® -therapy shows a quantitative

and qualitative increase of granulation tissue formation on the wound surface.

The mechanisms leading to this improved wound healing are still not known.

The increased microvessel density which is seen later most possibly reflects

the improved granulation formation and wound healing. Our hypothesis is

that V.A.C. ® -therapy promotes a better and accelerated wound healing by an

accumulation and activation of PMN at the wound site with an induction of

endothelial cell proliferation and angiogenesis. This accelerated angiogenesis

is promoted by mediators released from activated PMN.

We investigated the participation of neutrophil granulocytes in initialization

and propagation of the early wound inflammation and angiogenesis during

V.A.C. ® -therapy after trauma. Several parameters such as expression of integrins,

generation of oxygen radicals, release of cytokines and apoptosis are

analyzed in PMN and fluids isolated from the patient’s wound and compared

with serum and systemic PMN isolated from peripheral blood. The release of

different mediators was measured in ELISA.

From 58 patients serum and wound fluid samples (n=122) were obtained.

Cytokines and growth factors (IL-6, IL-8, IL-10, VEGF, FGF-2) were measured


y ELISA. Significantly higher levels for IL-8 and VEGF were observed in

woundfluids of VAC-treated patients with polytrauma, whereas other mediators

remained unchanged. The increased level of IL-8 and VEGF could

enhance inflammation and angiogenesis and thus might improved wound

healing.

Understanding the mechanisms involved in the wound healing during

V.A.C. ® -therapy might enable timely addition of healing-promoting factors

to the wound ground in the future. This could further improve the healing

processes, especially in infected wounds, or in wounds from severely injured

patients suffering from an essential immunosuppression during the posttraumatic

course.

Achievements 2005

� L. Labler, L. Mica, L. Härter, O. Trentz, M. Keel. Increased release of

Interleukin-8 and Vascular-endothelial growth factor in VAC-treated

wounds 4 th Day of Clinical Research, USZ, Zürich, 10-11 Mar 2005

� L. Mica, L. Härter, O. Trentz, M. Keel. NF-kB regulates LPS-induced

cytokine release, but NOT reduction of neutrophil apoptosis in patients

with sepsis 4 th Day of Clinical Research, USZ, Zürich, 10-11 Mar 2005

� M. Keel. New trend in osteosynthesis – The principle of angle stable

fixation.; Proximal humeral fractures – ORIF with LPHP – Locking

Proximal Humerus Plate.; Fractures of the femoral shaft – Antegrade with

AFN.; Fracture of the distal femur – LISS femur/LCP distal femur.; Tibial

plateau fracture – LISS and LCP tibia.; Limb salvage versus amputation.;

Computer assisted surgery in trauma care. AO International „Advances in

Fracture Management“;Hyderabad, India 31 Mar-03 Apr 2005

� Mica L, Härter L, Trentz O, Keel M. NF-kB reguliert die LPS-induzierte

Zytokinfreisetzung, nicht aber die Reduktion der Apoptose in neutrophilen

Granulozyten von Patienten mit Sepsis. Chirurgisches Forum 122. Kongress

der Deutschen Gesellschaft für Chirurgie, München, 05-08 Apr 2005

� Mica L, Härter L, Trentz O, Keel M. The IFN-γ-induced upregulation of

Toll-like receptors is regulated by STAT-3. 92. Jahreskongress der

Schweizerischen Gesellschaft für Chirurgie Zürich 8-10 Jun 2005

� Keel M, Labler L, Lustenberger T, Mica L, Stocker R, Trentz O. Outcome

after “Damage Control” or “Early Total Care” Management in 622

Severely Injured Patients. 92. Jahreskongress der Schweizerischen

Gesellschaft für Chirurgie Zürich 8-10 Jun 2005

� Keel M, Labler L, Lustenberger T, Mica L, Trentz O. Day-One-Surgery in 696

Severely Injured Patients by General Trauma Surgeons 92.

Jahreskongress der Schweizerischen Gesellschaft für Chirurgie Zürich

8-10 Jun 2005

� L. Labler. V.A.C. in der Traumatologie V.A.C. ® Drei-Länder-Kongress,

Graz, Österreich, 10-11 Jun 2005

� Pfiffner R , Labler L, Keller E, Pfammatter T. Erste Erfahrungen mit

einem neuen temporären Nitinol-Cavafilter. Schweizerische Gesellschaft

für Radiologie, Jahreskongress, Zürich, 16-18 Jun 2005


36

� M. Keel. Operationstaktik und Repositionstechniken bei Beckenfrakturen.

AO International, 4. Homburger Beckenkurs Teil II; Homburg, Deutschland

31 Aug-2 Sep 2005

� Mica L, Härter L, Trentz O, Keel M. STAT-3 reguliert die IFN-γ-induzierte

Expression der Toll-like Rezeptoren -2 und -4 in Leukozyten. 69.Jahrestagung

der Deutschen Gesellschaft für Unfallchirurgie, Berlin, 19-22 Okt 2005

� Labler L, Mica L, Härter L, Trentz O, Keel M. Erhöhte Interleukin-8 und

VEGF Spiegel in VAC-behandelten Wunden von Patienten nach Trauma.

69. Jahrestagung der Deutschen Gesellschaft für Unfallchirurgie, Berlin,

19-22 Okt 2005

� Keel M, Lustenberger T, Mica L, Lüthi S, Labler L, Trentz O. Der Schwere-

grad der Verletzungen und des haemorrhagischen Schocks korrelieren mit

der Inzidenz von Infektionen und septischen Komplikationen. 69. Jahrestag-

ung der Deutschen Gesellschaft für Unfallchirurgie, Berlin, 19-22 Okt 2005

� M. Keel. Das Querschnittssyndrom.Trauma-Management beyond ATLS ® ,

Zürich, 16 Nov 2005 (Keel Organisator).

� L. Labler. Diagnostik und Entscheidungsfindung beim SHT, Das Querschnitts-

syndrom.Trauma-Management beyond ATLS ® , Zürich, 16 Nov 2005

(Keel Organisator).

Collaborations:

� PD Dr. R. Graf, Division of Visceral & Transplant Surgery, USZ, Zürich

� Prof. Dr. D. Demitriades, Director of Trauma / Surgical Critical Care,

University of Southern California, Los Angeles, USA

Selected references:

� Keel M, Mica L, Stover J, Stocker R, Trentz O, Härter L. Thiopental-

induced apoptosis in lymphocytes is independent of CD95 activation.

Anesthesiology. 2005;103(3):576-584.

� Keel M, Trentz O. Pathophysiology of polytrauma. Injury. 2005;36(6):691-709.

� Bhandari M, Devereaux PJ, Tornetta P 3rd, Swiontkowski MF, Berry DJ,

Haidukewych G, Schemitsch EH, Hanson BP, Koval K, Dirschl D, Leece

P, Keel M, Petrisor B, Heetveld M, Guyatt GH. Operative management of

displaced femoral neck fractures in elderly patients. An international survey.

J Bone Joint Surg Am. 2005;87(9):2122-2130.

� Tschopp S, Keel M, Schmutz J, Maggiorini M. Abdominal compartment

syndrome after scuba diving. Intensive Care Med. 2005;31(11):1595.

� Ludwig Labler, Otmar Trentz, Marius Keel Traumatic Hemipelvectomy

European Journal of Trauma 2005;Vol 31(6):543-550

� Ludwig Labler, Jorn Zwingmann, Dieter Mayer, Reto Stocker, Otmar

Trentz, Marius Keel V.A.C. Abdominal Dressing System: A Temporary

Closure for Open Abdomen European Journal of Trauma; 2005 Vol

31(5):488-494

� Marius Keel, Ludwig Labler, Otmar Trentz „Damage Control“ in Severely

Injured Patients: Why, When, and How? European Journal of Trauma

2005;Vol 31(3):212–221


PD Dr. med.

Guido Wanner

Dr. med.

Peter

Richards,PhD

Dr. med.

Omana A.

Trentz

Sonja Hemmi

Dr. Marcus

Egermann

Dr. med.

Alexander E.

Handschin

Dr. med.

Markus Cardell

2.3.2 Osteogenesis Laboratory & Bone Research

PD Dr. med. G. Wanner, Dr. P. Richards, Dr. med. O. A. Trentz,

Dr. med. M. Egermann, Dr. med. A. E. Handschin, S. Hemmi

Fracture Healing in Osteoporosis

Osteoporosis, a major public health burden, is associated with increased fracture

risk. Fracture healing in osteoporosis is altered with reduced callus formation

and impaired biomechanical properties of newly formed bone leading to high

risk of fixation failure. Fracture healing in an established small animal model for

osteoporosis (SAMP-6 mice) is investigated using a newly developed angular

stable fixation device. Analysis is performed using histology, computed

tomography and biomechanical testing.

Bone marrow-derived mesenchymal stem cells (BMSC) provide an alternative

to bone grafting and composites enriched in BMSC demonstrated superior bone

healing. Matrices enriched in BMSC may increase the bone repair in osteoporosis-associated

fractures. Although BMSC from osteoporotic donors have

shown decreased proliferation and osteoblastic differentiation pattern in-vitro, it

is unclear whether BMSC from osteoporotic individuals possess similar healing

capacities in-vivo like BMSC from healthy donors. Furthermore it is unknown if

BMSC from osteoporotic donors could be stimulated to increase their proliferation

and differentiation pattern. In addition we study the influence of BMSC from

osteoporotic and healthy donors on bone healing.

Figure 1: Newly developed angular stable implant fi xing a midshaft osteotomy

in osteoporotic mice.


38

Effect of leptin on bone cells

Leptin, known as the obesity gene, is playing a crucial role in maintaining bone

turnover. In leptin knock out mice, the infusion of intraventricular recombinant

leptin into the hypothalamic area leads to a massive induction of osteoporosis.

In contrast to this central effect, there is also a hypothesis on a direct, peripheral

effect of leptin on bone cells. To further elucidate this hypothesis we

are currently investigating the effect of recombinant leptin on primary human

osteoblasts in vitro. One of our working hypothesis is that leptin may affect the

late mineralization stage of mature osteoblasts, enhancing the mineralization

of the extra cellular matrix.

Bone microtissue

Figure 2: Human osteoblast after 21 days of incubation under leptin infl uence

(left) and without (right).

During the last years, culturing of osteoblasts has been extensively studied

in this lab. The biochemical behaviour of cells attached to standard culture

devices doesn’t represent physiological processes in detail. Our approach of

bone tissue engineering includes a newly developed method of threedimensional

cell growth. This method lacks any scaffolds since they interfere

with cell metabolism. The planned studies, in collaboration with Professor

Hoerstrup, will examine the potential of osteoporotic BMSC to form mineralized

micro-tissue using the hanging drop technique. The ultimate goal of which is to

establish a multifunctional “microbone” in vitro which maybe used to enhance

our understanding of the mechanisms involved in bone disease.


Achievements 2005

� Egermann M: Present state and success of viral gene transfer in orthopaedics

with a focus on bone healing. EORS/EFFORT Symposium.

Lissabon June 4 th , 2005

� Egermann M: The influence of Ad-BMP-2 Gene Transfer on Bone healing

in osteoporosis. International Society of Molecular Orthopaedics.

Arosa April 2 nd , 2005

� Handschin A, Egermann M, Wanner G, Trentz O, Zund G, Trentz OA. Human

osteoblasts show depleted osteocalcin and Cbfa-1 (runx-2) expression in

heparin osteoporosis in vitro. SGC Zürich, 2005

Collaborations:

� AO Research Institute, Davos, Switzerland (Prof. K. Ito, Prof. M. Alini)

� AO Development Institute, Davos, Switzerland (R. Matthys)

� Departement of Clinical Research, University of Berne

(Prof. W. Hofstetter)

� Division of Regenerative Medicine, Department of Surgical Research,

University Hospital of Zurich (Prof. S. Hoerstrup, Dr. J. Kelm)

� PD Dr. med. HJ Kock, Unfallchirurgie, Hochtaunuskliniken Bad Homburg

Selected references:

� Egermann M, Goldhahn J, Schneider E: Animal models for fracture treat-

ment in osteoporosis. Osteoporos Int. 2005 Mar;16 Suppl 2:S129-38

� Egermann M, Schneider E, Evans CH, Baltzer AW: The potential of gene

therapy for fracture healing in osteoporosis.

Osteoporos Int. 2005 Mar;16 Suppl 2:S120-8.

� Trentz OA, Handschin A, Bestmann L, Hoerstrup S, Trentz OL, Platz A.

Influence of brain injury on early posttraumatic bone metabolism.

Crit Care Med. 2005 Feb;33(2):399-406.

� Handschin AE, Trentz OA, Hoerstrup S, Kock HJ, Wanner G, Trentz O.

Effect of low molecular weight heparin (dalteparin) and fondaparinux

(Arixtra) on human osteoblasts in vitro. Br J Surg. 2005 Feb;92(2):177-83.

� Handschin AE, Trentz O, Kock HJ, Wanner G. Low molecular weight heparin

induced skin necrosis. Langenbeck’s Arch Surg 2005 Jun;390(3):249-54

� Handschin AE, Egermann M, Trentz O, Wanner G, Kock HJ, Zund G,

Trentz OA. Cbfa-1 (Runx-2) and osteocalcin expression by human osteo-

blasts in heparin osteoporosis in vitro.

Clin Appl Thromb Hemost. Accepted Dec. 2005, in press.

� Handschin AE, Egermann M, Wedler V, Trentz O, Hemmi S, Trentz OA.

A comparative analysis of phenotype expression in human osteoblasts

from heterotopic ossification and normal bone.

Langenbeck’s Arch Surg. Accepted Dec 2005, in press.


40

PD Dr. med.

Michael

Heinzelmann

Dr. med.

Herbert Bosshart

2.3.3 Innate Immunity Laboratory

Targeting Bacterial Endotoxin

Michael Heinzelmann & Herbert Bosshart

The term sepsis describes a potentially lethal clinical condition that develops

as a result of a dysregulated host response to bacterial infection. The commonest

bacterial component implicated in initiating the septic syndrome is a cell wall

molecule derived from Gram-negative bacteria, known as lipopolysaccharide

(LPS) or endotoxin. Like all mammals, humans are equipped with an

LPS-sensing machinery consisting, primarily, of LPS-binding protein (LBP),

CD14, a glycosylphosphatidylinositol (GPI)-anchored monocyte differentiation

antigen, and Toll-like receptor 4 (TLR4), a signal transducing integral

membrane protein. Modest stimulation of TLR4 facilitates the elimination of

invading microorganisms. Potent TLR4 stimulation, however, produces severe

reactions in the host, often leading to multiple organ failure and death. The

search for pharmaceuticals that reduce mortality in septic patients has been

a painstaking process. Thus far, only a few compounds have been found to

significantly reduce mortality rates. Perhaps one of the more promising therapeutic

strategies currently pursued is based on the identification of synthetic

or naturally-occurring substances that neutralize LPS or inhibit LPS-mediated

activation of host immune cells such as monocytes and macrophages. Over

the past few years, we have identified a number of molecular structures with

a capacity to either enhance or blunt LPS-induced monocyte activation.

Our studies and those of others have uncovered several mechanisms by

which LPS-induced inflammatory responses in human whole blood are modified.

We showed that binding of LPS to cationic polypeptides can result in

both, enhancement or inhibition of LPS signals, depending on the type of

amino acid that contributes most to the cationic nature of the polypeptide.

Arginine- and lysine-rich cationic polypeptides are enhancers of LPS signals.

Histidine-rich polypeptides, in their protonated state, bind and neutralize LPS.

Cationic lipids act as LPS neutralizers by entrapment of LPS in micelles.

Sulfated polysaccharides like heparin or dextran sulfate associate with LBP,

displace LPS, and accelerate the transfer of LPS to other acceptor molecules,

e.g. soluble or membrane-bound CD14, resulting in an enhanced inflammatory

response of monocytes, and, presumably also of endothelial cells.

The molecular mechanisms of LPS enhancement and inhibition are of immediate

clinical relevance. The immunologically inert anticoagulant fondaparinux

can be used as a replacement for LPS-enhancing heparin, at least in patients

who are at risk for developing Gram-negative infections. Protamines, although

strong enhancers of LPS responses, are probably safe, as long as patients

are not treated with this polycation repeatedly. LPS-inhibiting histidine-rich

peptides, or cationic lipids are promising compounds for future therapeutic

interventions in patients with sepsis. However, for these and many other LPSneutralizing

substances, bridging the gap between bench and bedside, may

not be a straightforward process.


Models for the interaction of LPS-enhancing or -inhibiting substances with extracellular components of the

LPS activation pathway. (A) In human plasma, circulating LPS-LBP complexes are recognized by monocytic

CD14 (top vertical arrow). MD2-dependent activation of TLR4 initiates a signaling cascade that results in the

production and release of several proinfl ammatory mediators (bottom vertical arrow). Alternatively, LPS bound

to soluble CD14 activates endothelial cells which express TLR4 and MD2 but not CD14. (B) Cationic proteins,

e.g. protamines and CAP37, associate with LPS and bind to cellular surfaces. Subsequent steps (curved arrow)

leading to an enhanced LPS response (bottom vertical arrow) have not yet been addressed experimentally.

(C) Heparin exerts its anticoagulant effect through binding to AT III and thrombin. Apart from its interaction with

proteins of the coagulation pathway, heparin binds to LBP and displaces LPS (top vertical arrow) resulting in

enhanced LPS signals (bottom vertical arrow). (D) Poly-L-histidines (Hn) are devoid of stable secondary structures,

i.e. they exist as random coils. Similar concentrations of LBP and Hn result in near complete inhibition of

LPS responses in human whole blood (dashed arrows), presumably through adsorption of LPS to Hn (curved

arrow). (E) Cationic lipids neutralize LPS responses (dashed arrows) by incorporating LPS into micelles (curved

arrow).

Achievements 2005

Published Work

� Heinzelmann M, Bosshart H (2005). Heparin binds to lipopolysaccharide

(LPS)-binding protein, facilitates the transfer of LPS to CD14, and enhances

LPS-induced activation of peripheral blood monocytes. J Immunol, 174:

2280-7.

Lectures

� Bosshart H, Modulation of Host Responses to Bacterial Endotoxin.

International Conference on Environmental, Industrial and Applied

Microbiology, March 15 18, 2005, Badajoz, Spain.


42

Collaborations:

� Hans Flodgaard, Leukotech, Fruebjergvej 3, Box 8, 2100 Copenhagen -

Denmark

� Jean-Marc Herbert, Cardiovascular and Thrombosis Research

Department, Sanofi Synthelabo, 195 route d'Espagne , 31036 Toulouse

CEDEX - France

� Hans-Peter Beck, Swiss Tropical Institute, Socinstrasse 57, CH

4002 Basel - Switzerland

� Jerome Pugin, Geneva University Hospital Department of Internal

Medicine 24, rue Micheli-du-Crest, CH 1211 Geneva 14 - Switzerland

Selected references:

� Heinzelmann M, Bosshart H (2005). Heparin binds to lipopolysaccharide

(LPS)-binding protein, facilitates the transfer of LPS to CD14, and enhances

LPS-induced activation of peripheral blood monocytes. J Immunol, 174:

2280-7.

� Bosshart H, Heinzelmann M (2004). Lipopolysaccharide-mediated cell

activation without rapid mobilization of cytosolic free calcium. Mol

Immunol, 41:1023-8.

� Bosshart H, Heinzelmann M (2004). Human neutrophil-derived CAP37

inhibits lipopolysaccharide-induced activation in murine peritoneal

macrophages. Immunol Lett, 94:175-82.

� Heinzelmann M, Bosshart H (2004). Fondaparinux sodium lacks immu

nomodulatory effects of heparin. Am J Surg, 87:111-3.

� Bosshart H, Heinzelmann M (2003). Endotoxin-neutralizing effects of

histidine-rich peptides. FEBS Lett, 553:135-40.

� Bosshart H, Heinzelmann M (2002). Arginine-rich cationic polypeptides

amplify lipopolysaccharide-induced monocyte activation. Infect Immun,

70:6904-10.


PD Dr. med.

Dr. med.

2.3.4 Computer Assisted Trauma Surgery (CATS)

PD Dr. med. Peter Messmer, PD Dr. med. Guido A. Wanner,

PD Dr. med. Marius Keel, Dr. med. Felix Matthews, Dipl. Ing. FH Adrian John,

Dipl. Inf. Ing. ETH Adrian Egli, Dr. med., med. dent. Heinz-Theo Lübbers

Peter Messmer Felix Matthews

Computer assisted surgery has started with navigation about 15years ago in

Dr. med.

Valentin Neuhaus

Dipl. Inf. Ing. ETH

Adrian Egli

Dipl. Ing. FH

Adrian John

Dr. med., med. dent.

Heinz-Theo Lübbers

neurosurgery and soon later in ENT. PD Dr. Wanner and PD Dr. Keel introduced

2D navigation early in 2002 into our clinical work after a phase of

training and precision studies. With the liquidation of Medivision and transition

via Praxim to BrainLab there was a long period with no technical support for

laboratory and clinical projects in computer assisted surgery. Only in March

2005 the new fluoro navigation system of BrainLab became functional. So we

used the time to build a network of cooperations with the CARCAS Research

Group of the University Hospital of Basel, the AO Development Institute (ADI)

in Davos and some new industrial partners, such as Siemens Switzerland

and Siemens World (Erlangen, D), BrainLAB (Heimstetten, D) and Synthes

(Oberdorf, CH). Together with these partners we expanded our research

field from navigation to technology integration of different aspects and intraoperative

imaging in trauma. Siemens and BrainLab could be linked to our

university with contracts within different projects.

Together with AO-International (AOI), BrainLab and other partners from the

Computer-assisted Surgery Expert Group of the AO (University of Ulm, BG

Klinikum Ludwigshafen) teaching modules with workshops, videos and posters

were developed.

With a new software for navigated positioning of the fluoroscope precision

tests have been performed. They are under evaluation and preparation for

publication. Clinical tests will follow.

Another new software for navigated reduction control in maxillo-facial surgery

has been installed on the trauma navigation system and successfully been

used intra-operatively in 5 cases (Fig 1a+b) (Dr. Lübbers).

Figure 1a

Navigated control of correct reduction of the zygoma

fracture. Full arrow shows dynamic reference base

on the skull. Dotted yellow arrow shows referenced

pointer

Figure 1b

Display which shows the planning and the actual

positon of the bone after reduction.


44

Laboratory tests of stability of a passive holding device for computer aided

reduction and feasibility for intra-operative use of the device were published

this year (see publication record).

Together with Siemens, Synthes and CARCAS we are working on a software

solution (i3db) to improve workflow from admittance of the patient to preoperative

planning (fig 2), intra-operative handling, optimization of storage,

documentation, and accounting. The project involves different divisions of

the hospital. Meetings with OR personnel from several hospitals were hold.

A large KTI-grant application was accepted, so that we will employ a further

ETH - IT researcher in March 2006. This project is actually the most comprehensive

one of our research group.

Several other research grants have been written and accepted.

Figure 2

Virtual 3D planning and simulation

Achievements 2005

� Establishment and consolidation of cooperation with industrial partners:

Siemens, BrainLab, Synthes

� Cooperation with CARCAS Basel continued

� MaxFax navigation software of BrainLAB installed and used.

� MEPUC project: Precision testing performed

� I3db workflow project supported by a large KTI grant.

� Stability tests of passive holding device published.


Collaborations:

� PD Dr. Simon Wildermuth, Institut für Diagnostische Radiologie,

Universitätsspital Zürich

� Prof. Grätz, Dr. R. Eglmeier, Dr. H. Lübbers, Klinik und Poiklinik für

Kiefer- und Gesichtschirurgie, Universitätsspital Zürich

� Dr. D. Holzmann, Klinik für Ohren-, Nasen-, Hals- und Gesichtschirurgie,

Universitätsspital Zürich

� CARCAS, Research Group for Computer Assisted Radiology and Surgery

of the University Hospitals of Basel and Zurich

� Expert Group Computer Assisted Surgery of the AO Foundation with the

following partners:

- Prof. C. Krettek, Unfallchirurgische Klinik, Medizinische Hochschule

Hannover MHH

- Prof. F. Gebhard. Abteilung für Unfallchirurgie, Hand- und Wiederherstellungschirurgie,

Universitätsklinikum Ulm

- Prof. J. Alonso, Orthopaedic Trauma Service, University of Birmingham,

Alabama

- PD Dr. P.A. Grützner, Klinik für Unfall und Wiederherstellungschirurgie,

BG Unfallklinik Ludwigshafen

- PD Dr. U. Stöckle, Centrum für Muskuloskeletale Chirurgie, Charité,

Universtätsmedizin, Berlin

� AO Development Institut, Davos

� University of Applied Sciences Bern/Biel

� Siemens Med, Erlangen, Deutschland

� Siemens Schweiz, Zürich Altstetten

� BrainLab, Heimstetten, Deutschland

� Synthes, Solothurn

Selected references:

� T Gross, F Amsler, W Ummenhofer, M Zuercher, AL Jacob, P Messmer

and RW Huegli: Multiple-trauma management: Standardized evaluation

of the subjective experience of involved team members. European

Journal of Anaesthesiology, 22:754-761, 2005

� Felix Matthews, Valentin Neuhaus, Daniel Schmucki, Ronald Schwyn,

Thomas Gross, Pietro Regazzoni, Otmar Trentz, Peter Messmer:

Passive Pneumatic Stabilization Device for Assisting in Reduction of

Femoral Shaft Fractures. European Journal of Trauma, No 6, Vol 31,

568-574, December 2005

� Hügli RW, Staedele H, Messmer P, Regazzoni P, Steinbrich W, Gross T:

Displaced Anterior Column Acetabular Fracture : Closed Reduction and

Percutaneous CT-navigated Fixation. Acta Radiol. 2004 Oct. 45 (6): 618-21


46

PD Dr. med.

Guido Wanner

Ahmed Elsherbiny

Wiss. Mitarbeiter

Dr. med.

Claudio Contaldo

Andrea Schleh

Wiss. Mitarbeiterin

2.3.5 Musculosceletal tissue conditioning

Effects of recombinant Erythropoietin on critically perfused hamster

skin flap

Dr. med. Claudio Contaldo; Ahmed Elsherbiny, PD Dr. med. Guido A. Wanner

Originally, stimulation of erythrocytic progenitors was thought to be the only

physiological function of Erythropoietin (EPO). But EPO seems to have cytoprotective

actions as well as vascular functions. It has been reported that survival

of hypoxic tissue may be improved after EPO-treatment. The aim of the

present study was to assess the effect of systemically administered EPO on the

microhemodynamics in critically perfused skin flap tissue and to investigate the

mechanism of protection. The experiments were performed in the hamster

dorsal pedicled island flap model (Fig. 1), which consists of a proximal anatomically

perfused area, and a distal ischemic part (Fig. 2). By use of intravital

fluorescence microscopy the effects of intraperitoneally administred Recormon

(either 500 or 5000 IE/ kg bw) on the hypoxic microvasculature in the flap were

investigated up to 5 hours after onset of ischemia. To detect a possible mechanism

of action eNOS was blocked by L-NAME. In the ischemic flap area of control

(n=6) animals after 5 hours of ischemia, nutritive perfusion represented by the

functional capillary density (FCD) was decreased to 51 ± 2% (p< 0.01 vs.BL,

capillary diameters(CD) were dilated to 141 ± 2% (p< 0.01 vs.BL) and capillary

volumetric bloodflow (VBF)was diminished to 32 ± 5%(p< 0.001 vs.BL) of

baseline values. This microcirculatory dysfunction was substantially attenuated

by pretreatment with EPO 5000 IE/ kg bw (n=6); FCD 72 ± 8%, CD 119 ± 2%,

VBF 64 ± 8% of baseline (for all parameters p< 0.01 vs.Control). The lower

dose of EPO 500 IE/ kg bw showed an almost equal effectiveness (n=6); FCD

64 ± 9%, CD 122 ± 3%, VBF 50 ± 11% (for all parameters p< 0.01 vs.control,

and for VBF; p < 0.05 vs. EPO 5000 IE/ kg bw). The beneficial effects of EPO

were abolished when L-NAME was administred (n=6); FCD 53 ± 4%, CD 140 ±

3%, VBF 34 ± 6% (for all parameters p< 0.01 vs.control). In the same time we

observed a significant reduction of apoptotic cells in the dermal layer after EPO

treatment (Fig. 4) compared to control animals (Fig. 3). From our results we

conclude that Erythtropoietin may be used as a cytoprotective substance with

direct beneficial effects on the microvasculature in critically perfused tissue.,

which may be mediated by e-NOS. The fact, that the recombinant human form

of EPO is a widely used drug for the treatment of anaemia with a well-known

safety profile, makes it possible to use it soon in clinical trials.

Figure 1 Hamster flap Figure 2 Anatomic and ischemic part4


Figure 3 TUNEL positive cells in control skin Figure 4 TUNEL positive cells after EPO treatment

Achievements 2005

� Forschungspreis 2005 der Schweizerischen Gesellschaft für Plastische,

Rekonstruktive und Aesthetische Chirurgie (SGPRAC) für Dr. C.Contaldo

Talks 2005

� Erythropoietin-Doping in der elektiven Weichteilchirurgie?

Contaldo C, Trentz O, Menger MD, Wanner GA

122. Deutscher Chrirugenkongress (München, 05.04-08.04.05)

� Local heat preconditioning improves surival of critically ischemic porcine

flap, which may be mediated by induction of HO-1 and i-NOS

Contaldo C, Erni D. EURAPS (Marseilles, 26.05.-28.05.2005)

� EPO administration protects critically reperfused experimental osteomycu

taneous flap tissue. Contaldo C, Wanner GA, Künzi W

Schweizerische Gesellschaft für Plastische und Rekonstruktive und

Aesthetische Chirurgie (Biel am 30.9.05-01.10.2005)

Collaborations:

� Prof. Dr. MD Menger, Institut für Klinisch-Experimentelle Chirurgie,

Universitäts klinikum, Homburg/Saar, Deutschland

Selected references:

� Plock J, Contaldo C, von Ludinghausen M. Levator palpebrae superioris

muscle in human fetuses: anatomical findings and their clinical relevance.

Clin Anat. 2005

� Plock JA, Contaldo C, Erni D. Is hemoglobin in hemoglobin vesicles

infused for isovolemic hemodilution necessary to improve oxygenation in

critically ischemic hamster skin?

Am J Physiol Heart Circ Physiol. 2005

� Harder Y, Contaldo C, Erni D.Preconditioning with monophosphoryl lipid

A improves survival of critically ischemic tissue.

Anesth Analg. 2005


48

Dr. med.

Walter Kuenzi,

Director a.i. of

Plast. - Hand &

Reconstr. Surgery

Dr. med.

Volker Wedler

Manfred Welti

Prof. Dr. med.

Victor E. Meyer,

Dr. med.

Christian Köhler

2.4 Plastic, Hand & Reconstructive Surgery Research

Plastic, Hand &

Reconstructive

Surgery Research

2.4.1 Tissue Engineering

Tissue Engineering

Anti-Aging Therapy

The regenerative capacity of cartilage is known to be very poor and is therefore

a major problem in several diseases with severe cartilage loss - such

as rheumatoid arthritis - traumatic articular defects and cartilage defects

secondary to trauma or tumorsurgery. The reconstruction of a destroyed joint

has remained a difficult problem. The ability to reconstitute tissue structure

and function in vitro has tremendous clinical implications and is likely to

become very important in coming years. However, several problems remain.

One factor of major importance is the quality of the engineered cartilage and

the quality of the extracellular matrix in particular.

Engineering of articular cartilage using PEGT/PBT copolymer carriers

and autologous grafting to repair full-thickness defects in small joints

Dr. V. Wedler, Dr. Ch. Köhler, M. Welti

We have replaced a complete articular defect in the distal knee joint of twenty

White New Zealand Rabbits with autologous chondrocytes cultured on a synthetic

biodegradable scaffold. The project is a co-operation with Isotis Tissue

Engineering and extends for a time period of two years. The purpose of this

animal study is to determine the amount and quality of the engineered

cartilage, the incorporation of the

implant, degradation/replacement of

the scaffold and long term biomechanical

properties after 6 weeks, 3 and

6 months respectively. The implants

are evaluated with CT scans, histology,

DMMB assay, PCR, DNA analyse

and biomechanical testing.


Collaborations:

� IsoTis Tissue Engineering, Bilthoven, Netherlands

� Department of Materials, Institute of Polymers, ETH Zurich

� Department of Materials, Polymer Technology, ETH Zurich

� Institut für Labortierkunde, Universität Zürich, Irchel

Impact of x-rays on human chondrocytes in the area of tissue engineering

Dr. V. Wedler, Dr. Ch. Köhler, M. Welti

Tissue engineering of chondrocyte cultures in vitro permits the performing of

analyses of the metabolism as a function of the presence and absence of

different mediators. However, there are natural factors, which can have an

effect on human cell cultures. The present study investigates how cartilage

cells of human joints react to x-rays in quality and quantity. Special attention is

paid to the transillumination check at airports. Further, this study is to contain

an analysis regarding a possible calculation of the growth rate of chondrocytes

by means of a mathematical formula. The test showed that after irradiation of

the chondrocytes with differently large dose units, no signifi cant differences

occurred compared with the control group‘s values. This applies both to the

cultures exposed to luggage transillumination as well as for those exposed to

radiation devices of the department for radiology. Regarding the calculation of

cell growth, a mathematical formula could be established, with which a prediction

of the cell increase became possible. This corresponds to the fourth

derivative of a linear function. A direct acute cell damage affecting cell increase

or causing quality variances could not be observed. However, the question

about long-term damage of the cell nucleus‘ DNA remains and requires further

investigation. Due to our results the growth rate is predictable as a function of

time.


50

Optical differentiation between several scaffolds in tissue engineering

with human chondrocytes using light microscopy and SEM

Dr. Ch. Köhler, Dr. V. Wedler, M. Welti

At present, we are describing results in the tissue engineering of chondrocytes

from human cartilages by quantitative and qualitative measurements (MTT

and GAG). But some features are thereby not conclusively assessed, as for

instance: design, pattern of growth or cell cohorts. In this study, we reported

about various methods to show optical and photo-optical effects of four

different types of scaffolds. We analysed four types of scaffolds produced in

different institutes (Isotis/Netherland, Innovent/Germany and a new polymer

scaffold from China). Human articular chondrocytes were isolated after traumatic

amputation in the emergency room. The chondrocytes were expanded

in culture fl asks (either 75 cm 2 ) using DMEM with 10% FCS, gentamicin (50

μg/ml), and amphotericin B (Fungizone, 2.5 μg/ml). After 14, 28 and 56 days

we performed a light microscopy as well as a scanning electron microscopy

(SEM). Afterwards, we printed digital pictures for the measurement of growth

features. The evaluation has provided us with totally new visualisation and

has therefore given us new insights into the scaffold. Design and cohorts have

distinctive characteristics in the different scaffold materials. In this way we are

able to discover growth procedures. Light microscopy photography and SEM

are appropriate means to visualise the growth and functional design of cell

cultures in tissue engineering.

Preisträger BIONALE „Form follows Nature“ Wettbewerb der Deutschen Gesellschaft für Plastische Chirurgie

2005 München


2.4.2 Anti Aging

Anti-aging therapy in tissue engineering: An approach in growing human

chondrocytes

Dr. Ch. Köhler, Dr. V. Wedler, M. Welti

The advancement in medicine requires an ever-faster provision of biological

tissue substitutes. This also concerns the tissue engineering of cartilage cells.

In order to provide patients and physicians as fast as possible with autologous

substitutes in the case of degenerative illnesses or after trauma, it is necessary

to grow cells with the same quality and in the same quantity in a shortened

time frame. In this study the effect of a combination preparation with antioxidants

was tested with respect to an increase of the cell growth rate. We used

human cartilage cells. In the process the sterile fi ltered and solved substance

was added to the culture medium and a quantitative (MTT and GAG test) and

qualitative analysis (histology) conducted over the course of ten days. The

histological evaluation shows vital cells both in the vitamin and the control

group. Analysed over the course of ten days in the MTT, the chondrocytes

with vitamin additive reach the maximum quantity one day earlier and exceeds

the total number in cartilage cells in relation to the control group. The GAG

test corresponds to the results of normal chondrocyte cultures. The addition of

different growth factors permits growth acceleration of cartilage cells with the

same quality and quantity.

Anti Aging therapy- a new business for plastic surgeons?

Dr. Ch. Köhler, Dr. V. Wedler, M. Welti

Anti Aging therapy becomes increasingly important in many disciplines of medicine.

It is an old method in a trendy position. There are some other terms such

as better aging, rejuvenation or age medicine that all refer to the same concept.

But who works in this new fi eld of hormones, diets and sports therapy? There are

many institutes offering their services. In this study, we analysed actual treatments

in Anti Aging therapies and tried to draw a line between

scientifi c and commercial Anti Aging procedures. We drew a comparison of

actual promotions on several websites and in scientifi c publications on PUB-

MED. After that we described the main priorities of dermatology, plastic surgery,

gynaecology and other disciplines. A historical search of Anti Aging therapies

was analysed by several antiques

books (18th and 19th century). The

main business is controlled by the

cosmetics industry. Plastic surgery

is primarily concerned with operative

treatment (Botox, Liposuction etc).

Compared to this we fi nd abundant

scientifi c publications in all disciplines.

Looked at retrospectively, Anti

Aging has been established for a long

time. Anti Aging therapy should only

be performed by persons who know

what they are doing. An interdisciplinary

team is a decisive factor. It is important

to know the historical facts and

scientifi c consolidated fi ndings.


52

Collaborations:

� Dr. J.P. Hellermann, Departement für Kardiologie, Epidemiologie, USZ

� Prof. Dr. Th. Bächi Elektronenmikroskopisches Zentrum Zürich

� Klaus Marquard, Elektronenmikroskopisches Zentrum Zürich

� Dr. B. Bode, Departement Klinische Pathologie, USZ

� Prof. Dr. von Rechenberg, Tierspital Zürich

� Dr. J. Roos, Departement für Radiologie, USZ

� Dr. P. Neuenschwander, ETH Hönggerberg, Biomaterials

� Dr. M. Schnabelrauch, INNOVENT Jena

� Dr. Thomas Scholz, Departement Chirurgie USZ

� Dr. Ludwig Labler, Departement Chirurgie, USZ

� Dr. Sen, Departement of Orthopaedic Surgery. University of California,

San Francisco

� Mazda Farshad, Departement Chirurgie, USZ

Selected references:

� The value of the 3D gadolinium magnetic resonance angiography (MRA)

versus the conventional digital subtraction angiography (DSA) in the

presurgical planning of reconstructive surgeries in the area of the lower

extremities (under review)

C. Koehler, D. Weishaupt, W. Kuenzi, V. Wedler

� Anti-aging therapy in tissue engineering: An approach to growing human

chondrocytes (under review)

C. Koehler, M. Guggenheim ,J. P. Hellermann, B. Bode, V. Wedler

� Oropharyngeale Rekonstruktion mittels freiem Jejunumtransplantat nach

Tumor- und Stenosenresektion: Analyse von 53 Fällen (accepted 12/ 05

Hand- Plastische und Mikrochirurgie)

N. Krügel, C. Koehler, V. Wedler, W. Künzi

� Vacuum assisted closure: specific indications (under review)

C. Koehler, F.J. Jung, T. Scholz, L. Labler, A. Jandali, M. Comber, W.

Kuenzi, V. Wedler

� Anti-aging therapy in tissue engineering: An approach in growing human

chondrocytes (under review)

C. Koehler, J. P. Hellermann, B. Bode, M. Welti, V. Wedler

� Iatrogenic neurovascular entrapment injuries caused by reduction and

intramedullary fixation of fractures of the lower limb (accepted 12/ 2005

European Journal of Trauma)

V. Wedler, L. Labler, Ch. Köhler, M. Guggenheim, W. Künzi, O. Trentz

� Extensive hydrofluoric acid injuries: A serious problem

V. Wedler, M. Guggenheim, M. Moron, W. Künzi, VE Meyer

J Trauma, 58(4):852-7, 2005

� A comparative analysis of phenotype expression in human osteoblasts

from heterotopic ossification and normal bone

Handschin AE, Egermann M, Wedler V, Trentz O, Hemmi S, Trentz OA A

Langenbeck‘s Archives of Surgery, (accepted: 15.12.2005)


PD Dr. med.

Stephan Korom

PD Dr. med.

Stephan Korom

Dr. med.

Markus Cardell

Dr. med.

Wei Zhai

Dr. med.

Quiag Tan

Dr. med.

Sven Hillinger

Dr. med.

Sven Hillinger

Dr. med.

Ilhan Inci

Dr. med.

Stephan Arni

Prof. Dr. med.

Walter Weder

2.5 Thoracic Surgery Research

Thoracic Surgery

Research

2.5.1 Transplantation Immunology

Transplantation

Immunology

Tissue Engineering

Oncology

Immunotherapy

Clinical-related

research

Lung transplantation has become an effective therapeutic option in the treatment

of patients with end-stage pulmonary diseases. However, early acute

graft dysfunction continues to be a serious obstacle to successful lung transplantation,

accounting for significant postoperative morbidity and mortality. In

our established large and small animal models of unilateral lung transplantation

we investigated different substances in terms of early graft function

improvement.

Sildenafil extends Survival and Graft Function in a large Animal Lung

Transplantation Model

S.Hillinger, M.Cardell, W.Zhai, Q.Tan, S.Korom

Restoring intracellular cGMP and inducing NO-synthesis attenuates ischemiaassociated

early pulmonary allograft dysfunction. Phosphodiesterase-5

(PDE), predominantly expressed in lung tissue, plays a pivotal role in modulating

the cGMP/NO-synthase pathway in endothelial and epithelial cells. In this

study we evaluate the effect of employing sildenafil (Viagra ® ), a specific inhibitor

of PDE-5, to counteract ischemia/reperfusion (I/R) injury in a single lung

transplantation model of extended ischemia. Donor animals (weight matched

outbred pigs, 28-35kg) in the treatment group (I) (n=5) were injected with

0.7mg sildenafil/kg into the pulmonary artery (PA) prior to inflow occlusion.

For perfusion, Perfadex ® , containing 0.7mg sildenafil/l was used, and the

graft stored at 1 ºC in the perfusion solution. After 24 hour ischemia, unilateral

left lung transplantation was performed. Starting at reperfusion, group I

received continous sildenafil (0.7mg sildenafil/kg), over six hours. Except for

the sildenafil application, the control group (II) (n=4) was treated identically

(PGE 1 was injected into the PA). One hour after reperfusion, the right main

bronchus (MB) and right PA were occluded. Over the next five hours, cardiopulmonary

parameters (systemic aterial, PA, central venous, left atrial pressure,

pCO 2 , pO 2 ) were measured, including extravascular lung water (EVLW).

Thiobarbituric acid-reactive substance assay (TBARS) and myeloperoxidase

(MPO) analysis from lung tissue were run. All recipients of group I survived

the six hour reperfusion period, in contrast, all control animals died within

1-2 hours after occlusion of the right side.


54

In comparison to a marked rise in pulmonary vascular resistance (PVR) in

group II (>1000 dynes/sec/cm-5), PVR in group I remained stable, moderately

elevated from baseline (baseline: 150-180 dynes/sec/cm-5 vs. endpoint:

1000 dynes/sec/cm-5). EVLW in group I did not increase during reperfusion

(baseline: 6.75 ± 1.4mg/kg vs. endpoint: 6.7 ± 1.0mg/kg), in contrast to group

II, where pulmonary edema at two hours reperfusion preceded terminal graft

failure (group I: 6.48 ± 1.8mg/kg vs. group II: 9.7 ± 0.1mg/kg). Tissue reactive

free radicals at endpoint measurement in group I did not differ significantly

from native tissue, yet, when compared to specimen taken from group II at

time of terminal graft failure, a significant increase in free radicals was noted

(group I: 13.8 ± 1.6 pmol/g vs. group II: 18.5 ± 3.0 pmol/g, p < 0.05).

Sildenafil treatment prevents terminal early graft failure, allowing lung transplantation

after 24 hours ischemia time. Reperfusion edema was strikingly

diminished, preserving pulmonary structural integrity and functional while

prolonging graft ischemia time. Employing the established PDE-5 inhibitor

sildenafil during lung perfusion, storage and implantation, ischemic tolerance

may be extended and early graft function improved.


Circadian Melatonin Secretion During Alloantigen Challenge and

Immunosuppression

M.Cardell, W.Zhai, FJ.Jung, S.Hillinger, S.Korom

Melatonin (MLT) displays a dose-dependent immunoregulatory effect, where

high-dose therapy has been shown to abrogate acute rejection and signifi -

cantly prolong graft survival. Endogenous melatonin secretion, in response to

heterotopic rat cardiac allograft transplantation, was investigated during the

acute rejection phase and under standardized immunosuppessive maintainance

therapy with CsA and rapamycin.

Cardiac grafts were harvested and transplanted to the abdominal great vessels.

Recipients (n=6/groups) of syngeneic transplants (LEW to LEW, group I), untreated

recipients of allogeneic grafts (LBNF1 to LEW, II), and allografted,

CsA- (III) or rapamycin-treated (IV) hosts constituted the experimental groups.

Endogenous circadian melatonin secretion was measured with a novel species

specific RIA for MLT, without alteration of the light/dark cycle in recipients

following transplantation, and correlated with the ensuing acute rejection response.

Furthermore, the infl uence of therapeutical immunosuppressive regimens

with CsA (2,5mg/kg bw) and rapamycin (3mg/kg bw) on circulating

melatonin levels over time was analyzed.

Neither the operative procedure alone, nor the allogeneic challenge with a

perfused allograft undergoing acute rejection infl uenced the endogenous melatonin

secretion pattern could. Whereas immunosuppression by CsA did not

affect circulating melatoni kinetics, rapamycin therapy signifi cantly lowered the

circadian secretory amplitude of endogenous melatonin.

Taken together, endogenous melatonin secretion is a stable mechanism. The

natural course of circadian endogenous melatonin secretion displays a robust

mechanism, which is unaltered by perfused allograft transplantation.


56

CD26/Dipeptidyl-Peptidase IV (DPP IV) Inhibition Attenuates Posttransplant

Pulmonary Ischemia/Reperfusion Injury after Extended Ischemia

W.Zhai, M.Cardell, I.Inci, S.Arni, S.Korom

The T cell costimulatory Ag CD26 possesses dipeptidyl peptidase IV (DPP

IV) catalytic activity, which is linked to its costimulatory effi cacy. Based on our

observation that abrogation of acute pulmonary rejection and preservation of

ventilatory function can be achieved by DPP IV inhibition in an rat allogeneic

lung transplantation model, to elucidate the non-immunological, enzymaticactivity-associated

effect of CD26/DPP IV following transplantation, in this

study we investigated the impact of specifi c catalytic inhibition of CD26/DPP

IV on ischemia-reperfusion (I/R) injury. A syngeneic rat orthotopic left lung

transplantation model was used. Group I animals consisted of control group,

donor lungs were fl ushed and preserved in Perfadex for 18 hours at 4 C and

then transplanted and reperfused for 2 hours; Group II animals received the

same procedures as group I except 25umol/L DPP IV inhibitor in both fl ush

and storage solutions; Group III animals received the same procedures as

group I except 25umol/L DPP IV inhibitor in fl ush solution only (6 Tx/group).

After 2-hour reperfusion, blood gas analysis and peak airway pressure (PAwP)

were measured, lung tissue biopsy specimens were obtained for assessment

of wet/dry weight ratio, myeloperoxidase activity (MPO) and thiobarbituric

acid reactive substances (TBARS).In comparison with control group, two DPP

IV inhibitor treated groups signifi cantly improved postreperfusion graft pO2,

PAwP, wet/dry weight ratio and lipid peroxidation (p < 0.05, respectively). In

this experimental model, posttransplant lung ischemia reperfusion injury can

be attenuated by selectively targeting graft DPP IV enzymatic activity.

Graft oxygenation at the end of

2h reperfusion

Peak airway pressures (PawP)

in mmHg during 2-hour reperfusion

period


Effect of N-Acetylcysteine on lung ischemia-reperfusion injury in rat

single lung transplantation

I.Inci, S.Arni, W.Zhai, M.Cardell

Several studies have shown that agents such as prostaglandins; the oxygen

free radical scavengers superoxide dysmutase, catalase, glutathione, allopurinol,

dimethylthiourea, lazaroids, trimetazidine; aprotinin; platelet factor antagonists;

and angiotensin converting enzyme inhibitor, captopril and melatonin to

be effective in protecting lung against ischemia-reperfusion injury. N-Acetylcysteine

(NAC) is a precursor of the most important physiological antioxidant

glutathione. Sulphydril-containing compounds, especially reduced glutathione

(GSH), are important in the protection of cells against hydroperoxide damage.

This important reducing agent and antioxidant is involved in maintaining the

cellular oxidation-reduction balance, and has been shown to protect cells from

a wide variety of endogenous and exogenous insults. GSH can also scavenge

free radicals produced by oxidative challenges.There have, therefore, been

many suggestions that reduced GSH may be useful therapeutically as an

antioxidant and cytoprotective agent. In this experimental study we wanted to

investigate whether donor and recipient treatment with NAC would reduce

ischemia-reperfusion injury following lung transplantation after 18 hours of cold

ischemic storage in a rat single lung transplant model. Orthotopic single left

lung transplantation was performed in male Fischer (F344) rats weighing 280-

300 g, using a cuff technique for the anastomoses. Animals were randomized

into two groups (n=5, each); ischemic control (IC) (group I) group; 18 hours

cold (4°C) ischemia followed by transplantation, intraperitoneal saline injection

(1 ml) 15 minutes before harvest and reperfusion, respectively, no treatment;

NAC treated (group II) group; transplantation was carried out after 18 hours of

cold ischemia (4°C), donor and recipient treatment with intraperitoneal injection

of 150 mg/kg NAC 15 minutes prior to harvest and reperfusion, respectively.

Right donor lungs (n=5) were assessed for GSH, MPO and TBARS in order to

obtain baseline values in normal lung. Oxygenation 2 hours after graft reperfusion

was higher in the NAC group (184.5±83.3 mmHg) than in the IC group

(67.3 ± 16.4 mmHg) (p=0.016). Peak airway pressure (PawP) measurements

made during the reperfusion period showed signifi cant differences among the

groups (p=0.015). At the end of 2-hour reperfusion PawP was 14.4±1.6 cm

H2O in NAC group, and 19.2±2.2 cmH2O in IC group (p=0.008) (Fig. 1). The

amount of lipid peroxidation was signifi cantly higher in IC (17.46±10.6 micromol/g)

compared to NAC (7.34±1.9 micromol/g) group (p=0.016). Reduced

glutathione (GSH) level in the normal lung was 37.6±5.4 μM. GSH levels in

IC and NAC groups were 6.8±0.9 μM and 20.6±2.4 μM, respectively. The

difference between the groups was statistically signifi cant (p=0.004) (Fig. 2).

Myeloperoxidase activity (MPO) in IC and NAC groups were 0.52±0.44 units/

g, 0.34±0.4 units/g, respectively. The difference between the groups was not

signifi cant (p=0.3). W/D weight ratio in IC was 8.2±105 and 6.7±1.6 in NAC

group. The ratio was not different statistically between the two groups (p=0.2).

In this model, we have shown that recipient and donor treatment with NAC

protected lungs from reperfusion injury after prolonged ischemia. This effect

could be due to NAC’s free radical scavenging activity.


58

Fig 1. Peak airway pressures (PawP) in cm H2O during the 2-hour reperfusion period. The analysis of variance

for repeated measures using all measurements made during the reperfusion period differed signifi cantly

between the IC and NAC groups (p=0.015). At the end of 2-hour reperfusion , PawP was signifi cantly less in

the NAC group than in the IC group (p=0.008). (Int= intubation; Thor= when the thorax is opened; prerep = just

before reperfusion)

Fig. 2. Lung tissue reduced glutathione (GSH) levels between the groups showed a better preservation of GSH

in NAC group compared to IC group (p=0.004).

Achievements 2005

� M.Cardell was awarded a grant for the Sildenafil project from the

Hartmann-Müller foundation

� Sildenafil extends survival and graft function in a large animal lung trans-

plantation model

S.Korom, S.Hillinger, M.Cardell, W.Zhai, Q.Tan, W.Weder,

EACTS/ESTS Annual Meeting, Barcelona, Spain, Sept.2005, paper in

press

� Melatonin, CD26, NAC manuscripts in preparation

� M.Cardell was teaching orthotopic rat lung Tx in the microsurgery course

of the Division of Surgical Research Oct. 2005


Collaborations:

� Drs. Manz und Welp, Labor Diagnostika Nord GmbH & Co KG, Nordhorn,

Deutschland

� Dr. I. De Meester & Prof. Dr. S. Scharpé from the Dept. of Pharmaceutical

Sciences, University of Antwerp, Antwerp, Belgium

� PD Dr. M. Keel and Dr. L. Härter, Dept. of Traumatology, USZ, Zurich

Selected references:

� F. J. Jung , L. Yang , L. Härter, I. Inci, D. Schneiter, D. Lardinois, M. Keel,

W. Weder S. Korom. Melatonin in vivo prolongs cardiac allograft survival

in rats. Journal of Pineal Research, 37: 36-41 (2004).

� Korom S, De Meester I, Stadlbauer THW, et al. 1997. Inhibition of CD26/

dipeptidyl peptidase IV activity in vivo prolongs cardiac allograft survival

in rat recipients. Transplantation 63:1495-1500.

� De Meester I, Korom S, Van Damme J, and Scharpé S. 1999. CD26, let it

cut or cut it down. Immunology Today 20: 367-375.

� Hosepund JD, Bennet LE ,Keck BM, Fiol B, Boucek MM, Novick RJ. The

registry of the International Society for Heart and Lung Transplantation:

sixteenth official report- 1999. J Heart Lung Transplant 1999;18:611-26.

� Inci I, Dutly A, Rousson V, Boehler A, Weder W. Trimetazidine protects the

energy status after ischemia and reduces reperfusion injury in a rat single-

lung transplant model. J Thorac Cardiovasc Surg 001;122(6):1155-1161.

� Konukoglu D, Cetinkale O, Bulan R. Effects of N-Acetylcysteine on lung

glutathione levels in rats after burn injury. Burns 1997;23:541-44.

� Nakano H, Boudjema K, Alexandre E, et al. Protective effects of N-

Acetylcysteine on hypothermic ischemia-reperfusion injury of rat liver.

Hepatology 1995;22:539-45.


60

Dr. med.

Qiang Tan

Manfred Welti

2.5.2 Tissue Engineering

Novel aspects of tissue engineered trachea

Q.Tan, M.Welti

Our research focuses on the reepithelialization of tissue engineered trachea.

To achieve this goal, we have to overcome three main obstacles:

1. Identify epithelial cells source

2. Optimize epithelial cells seeding method

3. Expediate revascularization process

Regarding the source of epithelial cells, our hypothesis is that it is not the kind

of epithelial cells but the intactness of the basement membrane which plays

a key role in the reepithelialization of tissue engineered trachea. Many

researchers around the world have proved that obtaining enough tracheal

epithelial cells through autologous biopsy is very difficult. Therefore in our

study we plan to use skin keratinocytes, or even skin fleet, instead of tracheal

epithelial cells as the epithelial source. Fortunately, in the field of tissue

engineering, tissue engineered skin remains the most successful branch so

far. We plan to reconstruct tissue engineered skin first in vitro then used it

to cover the inner surface of tracheal scaffold. In the former exam we have

established the porcine epithelial cell culture protocol and touched upon the

reconstruction of tissue engineered skin.

With regard to cell seeding process, many earlier papers suggest that the

direct epithelial cell fleet seeding method works more efficiently than the cell

suspension seeding alternative. Accordingly, we decide to directly adhere

tissue engineered skin or fresh autologous thinner skin fleet to the scaffold

with the help of stent which has been already widely used to cure benign

tracheal stenosis in clinic.

Finally, revascularization remains to be the most difficult. Traditionally, tissue

engineered trachea is pre-implanted into the omentum and then transplanted

with pedicel blood vessels as a tracheal replacement. This, however, is proven

to be a complex process that entails laparotomy twice, with high risk of

complication. We therefore bring forward a novel concept of “in-vivo bioreactor”

defined as the design of a perfusion system inside the scaffold.The device

consists of two parts: an extracorporeal pump system and an intracorporeal

perfusion tube inserted into the tissue engineered trachea scaffold. The idea

is simple: the extracorporeal pumps will continuously administrate medium

through the porous perfusion tube into the tissue engineered trachea to sustain

the seed cells before a functional capillary net takes shape from normal

tissues around. The process resembles the principle of heart-lung machines

to maintain the normal blood circulation during heart operations. Through

this perfusion system fresh seed cells can also be added to refresh the seed

cells. This offers a clear advantage in emergency treatment when we have

not enough time waiting for the cell culture. Apparently we can also deliver

various growth factors to facilitate seed cells differentiation or accelerate the

angiogenesis process. All the above three possibilities have already been

proved in previous in vitro researches in our lab. Since we cannot accurately

mimic in vitro the entire in-vivo environment, a problem constantly bothering

researchers on bioreactors, it makes sense to treat the recipient himself as a

bioreactor. Therefore we name the design “in-vivo bioreactor”.


Fig. TE1. sketch of tissue engineered trachea with in-vivo bioreactor design.

A simple test model was established by inserting a porous catheter which

connected to a pump system into a tubular Degrapol scaffold. Tests were performed

on the advantages of maintaining the survival of epithelial cells seeded

onto the DegraPol surface and of effectively delivering chondrocytes into the

scaffold. Furthermore, based on chorioallantoic membrane CAM angiogenesis

model, we judged its effect on the scaffold angiogenesis process. Scanning

electronic microscopy SEM results show living epithelial cells after two weeks’

support by in-vivo bioreactor. In cell delivery test, MTT results proved no signifi

cant difference between in-vivo bioreactor delivery group (0.312±0.177) and

directly seeding static culture group (0.288±0.053). Regarding angiogenesis

test, in-vivo bioreactor groups presented more tissue in-growth and capillary

formation. These results demonstrated that through in-vivo bioreactor, we can

deliver and maintain the survival of seeded cells as well as accelerate the

scaffold angiogenesis process.

Fig TE2: SEM result show 16HBE14o cells adhered well to the DegraPol scaffold surface and survive after 2

weeks supported by in-vivo bioreactor.


62

Fig TE3: A: Histological slice from in-vivo bioreactor

group showed larger chondrocyte cluster formation.

B: In comparison, there is only a monolayer of chondrocytes

covered the surface of DegraPol tube in

directly seeding static culture group.

(hematoxylin and eosin staining, ×100)

Fig TE5: A: In the IV group erythrocytes migrate

throughout the whole DegraPol scaffold, even at the

upper part of the Degrapol tube without any other ingrowth

connective tissues, due to increased vessel

permeability caused by VEGF. (hematoxylin and

eosin staining, ×200) B: Bisbenzimide H 33342

staining marked all the erythrocyte nuclear red, proved

they were migrating from normal function vessels.

Achievements 2005

� Grant EMDO-Stiftung, manuscripts in preparation

Collaborations:

� Dr. P. Neuenschwander, Institute of Polymer Research, ETH, Zurich,

Switzerland.

� Dr. L. Moroni, Twente University, IsoTis S.A., Bilthoven, The Netherlands.

� Tissue Engineering Groups, Division of Surgical Research, USZ, Zurich.

� Dr. R.Steiner, Onkologie, USZ

Selected references:

Fig TE4: Scales of tissue in-growth in angiogenesis

test. A: grade 0, no tissue in-grow B: grade 1, CAM

tissue invades less than 1/3 part of the DegraPol

tube C: grade 2, tissue invasion more than 1/3 less

than 2/3 part of the DegraPol tube D: grade 3, CAM

tissue invasion include the whole DegraPol tube

(hematoxylin and eosin staining, ×200)

Fig TE6: Bisbenzimide H 33342 staining presents

normal functional capillary formation in Degrapol

scaffold in-growth tissue. The dye was injected into

CAM vein far away form the DegraPol scaffold and

marked the nuclear of all the cells which irrigated by

normal circulation system red.

� Yang L, Korom S, Welti M, Hoerstrup SP, Zund G, Jung FJ,

Neuenschwander P, Weder W.

Tissue engineered cartilage generated from human trachea using DegraPol

scaffold. Eur J Cardiothorac Surg. 2003 Aug; 24(2): 201-7.


Dr. med.

Sven Hillinger

Dr. Dr. med. med.

Isabelle Isabelle Opitz Opitz

Manfred Welti

Dr. med.

Stephan Arni

Dr. med.

Markus Cardell

Dr. med.

Didier Lardinois

2.5.3 Oncology

Loco-regional chemokine treatment inhibits tumor growth in an

orthotopic model of lung cancer

S. Hillinger, M.Cardell, S.Arni

Effective anti-tumor responses require both antigen presenting cells and

lymphocyte effectors. Although lung cancers express tumor antigens, they

are ineffective as antigen presenting cells because tumor cells often have

limited expression of MHC Ags and lack co-stimulatory molecules. It has

been demonstrated that local and systemic administration of chemokines as

stimulators of the immune response is beneficial as potent anti-cancer

strategy. Epstein Barr virus-induced molecule 1 ligand chemokine (ELC/

CCL19) is a CC chemokine that strongly chemoattracts both dendritic cells

(DC) and T lymphocytes. In this study we evaluated the anti-tumor efficacy of

loco-regional ELC/CCL19 administration in an orthotopic model of bronchogenic

carcinoma.

5x10 4 L1C2 and 3LL cells have been injected into the right upper pulmonary

lobe of Balb/C and C57/Bl6 mice respectively. Two days after injection mice

have been treated with right intraaxillar (lymph node region) injection of

recombinant ELC/CCL19 (0.5μg/dose) three times per week for 2 weeks. For

the evaluation of ELC/CCL19 mediated loco-regional anti-tumor responses,

lungs were harvested three weeks after treatment and lung blocks as well as

axillary lymph nodes were assessed for H&E staining to evaluate the tumor

burden and analyses of tumor infiltrating T cell subsets by flow cytometry.

Tumor bearing lungs were evaluated for the production of IL-10, IL-12, GM-

CSF, IFNγ, TGFβ, by ELISA and PGE2 by enzyme immunoassay (EIA) in the

supernatants after an overnight culture.

Histological evaluation of tumor sections and axillary lymph nodes revealed

extensive lymphocytic infiltration with a marked reduction in tumor growth

compared to diluent controls. Flow cytometric analysis showed a significant

increase in both CD4 and CD8 subsets as well as dendritic cells. However,

there was a decrease in CD4+CD25+ T regulatory cells in the tumor infiltrating

lymphocytes of ELC/CCL19 treated mice lungs. Lung tissue cytokine

profiles showed a shift towards immunostimulatory molecules.

These results in a clinically relevant orthotopic model of lung cancer confirm

the importance of chemokines in the development of an effective anticancerimmunotherapy.

Further studies are warranted to delineate the mechanisms

responsible for the anti-tumor responses following ELC/CCL19 therapy for

lung cancer.

Under general anesthesia a small skin incision and muscle cut is performed.

5x10 4 tumor cells are injected under vision of the moving lung into the right

upper pulmonary lobe. Inzision is closed with 1-2 single sutures.


64

Furthermore we have achieved fi rst promising results of a combination treatment

with the chemokine ELC/CCL19 and Interleukin-7, which amplifi es the

longevity of the tumor antigen specifi c T cells and NK effectors.

Achievements 2005

� Grants Zürich Cancer League, Sassella-Stiftung

� Presentations at the Annual Meeting of the AACR, Anaheim, Ca,

April 2005, manuscripts submitted and in preparation, several grant

applications running

Collaborations:

� Prof. S.M. Dubinett, Director of the UCLA Lung Cancer Program, and

Dr. S. Sharma, Associate Research Professor, University of California

Los Angeles

Selected references:

� Hillinger S, Yang SC, Zhu L, Huang M, Duckett R, Atianzar K, Batra RK,

Strieter RM, Dubinett SM, Sharma S. Epstein Barr Virus-Induced Molecule 1

Ligand Chemokine (ELC/CCL19) Promotes IFNg-dependent Antitumor

Responses in a Lung Cancer Model. J Immunol. Dec 15;171 (12):6457-

65 (2003).

� Yang SC, Hillinger S, Riedl K, Zhang L, Zhu L, Huang M, Atianzar K, Kuo BY,

Gardner B, Batra RK, Strieter RM, Dubinett SM, Sharma S. Intratumoral

administration of dendritic cells overexpressing CCL21 generates systemic

antitumor responses and confers tumor immunity. Clin Cancer Res. Apr

15;10 (8):2891-901 (2004)


Malignant pleural mesothelioma – investigation of different cytotoxic

substances

I.Opitz, S.Hillinger, D.Lardinois, S.Arni

Objective: To evaluate the cytotoxicity induced by taurolidine and povidoneiodine

(PVP-I) in malignant mesothelioma cells.

Materials and Methods: MTT cell viability tests were performed on four

human mesothelioma cell lines (ZL 5, ZL55, ZL34, NCI-H28) and on human

fibroblasts after exposure to taurolidine and PVP-I. Caspase 3-like assays,

flow cytometry with Annexin V and 7AAD, and Hoechst nuclear staining were

performed to assess the type of cell death induced by these agents. The

effect of taurolidine or povidone-iodine on p53 activation was measured by

Western blotting and their ability to sensitize cells to cisplatin was determined

in MTT- and caspase assays.

Results: Both substances killed mesothelioma cells already after 7.5 min

incubation, but taurolidine was more specific towards tumour cells. Taurolidine

but not PVP-I induced caspase-3-like activity in mesothelioma cell lines. Cell

growth inhibition by taurolidine could be reduced by the caspase inhibitor

zVADfmk. Annexin V/7AAD double labeling and Hoechst nuclear staining

revealed that taurolidine induced both apoptosis and necrosis after 24 h,

whereas PVP-I induced uniquely necrosis. Taurolidine activated p53 in mesothelioma

cells and sensitized them to cisplatin-induced apoptosis whereas

povidone-iodine had no such effect.

Conclusion: Both substances are cytotoxic to human malignant mesothelioma

cells at early and late time points at concentrations used in clinical

practice. Taurolidine induces apoptosis and necrosis in mesothelioma cells,

activates p53 and sensitizes cells to cisplatin, whereas PVP-I inhibits cell

growth via necrosis. Both solutions might be promising candidates for local

treatment concepts for malignant pleural mesothelioma.

Furthermore we started to investigate the intrapleural use of both substances

in vivo. In a first step we were able to establish the first recurrence model of

malignant mesothelioma in the rat. By subpleural injection of tumour cells

we observed a defined tumour growth of a reproducible size leading to local

recurrence exactly at the same site after resection of the nodule. With this

model we started now to investigate the influence of the intrapleural use after

extrapleural pneumonectomy of taurolidine, PVP-I, CCL-19, a chemokine that

influences cell mediated immune-respone at the tumour site in lung cancer

therapy. Furthermore the effect of intrapleural use of cisplatin solution or

cisplatin combined with a surgical sealant was observed.

Western blot after incubation of human malignant

mesothelioma cell line ZL55 with taurolidine

0.002% and 0.004% and povidoneiodine

0.05% and 0.01% for 24 hours


66

IC50 of povidone-iodine

and taurolidine after incubation

of different human

meosthelioma cell lines

and fi broblasts

Achievements 2005

� Zürich Cancer League „New treatment approaches for local control in

malignant pleural mesothelioma”

Collaborations:

� Laboratory for Molecular Oncology (Dr. phil. S. Hopkins-Donaldson and

Prof. Dr. R. Stahel)

� Department of Clinical Pathology (Dr. med. P. Vogt)

� Department of Biostatistics (Dr. V. Rousson)

Selected references:

ZL55

Povidone-iodine (%)

ZL55

Taurolidine (%)

� Rusch V., Niedzwiecki D., Tao Y., Markman M. Intrapleural cisplatin and

mitomycin for malignant mesothelioma following pleurectomy: pharmaco-

kinetic studies. J Clin Oncol 1992; 10: 1001 6.

� Opitz I, Van Der Veen HC, Braumann C, Ablassmaier B, Fuhrer K, Jacobi

CA. The influence of adhesion prophylactic substances and taurolidine/

heparin on local recurrence and intraperitoneal tumor growth after

laparoscopic-assisted bowel resection of colon carcinoma in a rat model.

Surg Endosc. 2003 Jul;17(7):1098-104. Epub 2003 Apr 28.

� Docherty JG, McGregor JR, Purdie CA, Galloway DJ, O´Dwyer PJ.

Efficacy of tumoricidal agents in vitro and in vivo. Br J Surg 1995; 82:

1050-1052


PD PD Dr. Dr. med.

med.

John John F. F. Stover

Stover

Silke Silke Ludwig Ludwig

Prof. Prof. Prof. Dr.

Dr.

Reto Reto Reto Stocker

Stocker

Jutta Sommerfeld

Sommerfeld

2.6. Surgical Intensive Care Medicine

The Division of Surgical Intensive Care Medicine consists of three Intensive

Care Units (ICU) in which critically ill surgical and medical patients are

treated. These patients suffer from various complex and complication- ridden

diseases, which require intense treatment of the underlying diseases (as

e.g., traumatic brain injury, pulmonary, hepatic, renal illnesses, burns) and

secondary life-threatening complications (as e.g., ARDS, sepsis, multi-organ

failure). Research within this field is extremely challenging as progressive

pathophysiologic changes are overshadowed by evolving secondary processes

and influenced by therapeutic interventions. The main aims of our

research employing analytical techniques in the newly established laboratory

are to 1) characterize important pathophysiologic and pharmacodynamic

cascades and 2) unmask potentially unfavorable effects of routine therapeutic

measures.

Our research focuses on disease- specific as well as disease- spanning

changes.

A) Disease- specific changes

In regard to disease- specific changes we are investigating the impact of

various brain lesions following severe traumatic brain injury on arterio- jugular

venous differences in amino acids, ATP degradation products, and different

cytokines which might differentiate lesion- dependent mediators from markers

of cell damage and unmask a pathophysiologic important temporal profile

possibly driving ensuing deterioration.

B) Disease- spanning changes

Nutritional requirements. In regard to disease- spanning changes we are

investigating cellular and humoral alterations with the aims of improving

nutritional requirements and reducing cellular immunodepression. This also

includes assessing the influence of different enteral and parenteral nutritional

solutions on potentially harmful amino acids as e.g., glutamate, arginine, and

aromatic amino acids.

Effects of norepinephrine. Norepinephrine is routinely used to elevate arterial

blood pressure with the aim of improving microcirculation and tissue oxygenation.

However, norepinephrine may compromise this aim by excessive

arteriolar vasoconstriction and activation of thrombocytes which could induce

microthrombosis, thereby promoting tissue injury. In addition, norepinephrine

might impair the activity of neutrophils, thereby contributing to cellular immunodepression,

thus predisposing the critically ill patient to the development of

infections. In this context, we have established the evaluation of sublingual

in vivo microscopy using the novel bed side and non invasive orthogonal

polarized spectral (OPS) imaging in critically ill patients which revealed a

dose- dependent norepinephrine- mediated decrease in vessel diameter and

functional capillary density (fig. 1).


68

Fig. 1 Infl uence of norepinephrine on sublingual microcirculation determined by OPS imaging in a healthy

control (left) and a patient (right) receiving high dose norepinephrine at 15 μg/ min. The functional capillary density

was signifi cantly decreased from 47.3 to 28.8 cm/ cm 2 . (Doctoral thesis of Claudine Fridez)

In vitro stimulation of isolated platelets and neutrophils revealed a significant

concentration- dependent increase in platelet activation (fig. 2) and neutrophil

impairment (fig. 3), respectively.

Surveillance of pharmacological and metabolic coma. Depending on the

underlying disease, continuous administration of analgetics and sedatives

becomes indispensable. However, individual pharmacogenetics, elimination

and distribution profiles might predispose the patients to iatrogenic induced

drug tolerance and dependency with ensuing severe withdrawal symptoms

due to cerebral transmitter system imbalance.

Fig. 2

Dose- dependent increase in surface P- Selectin

expression in isolated thrombocytes from healthy

controls stimulated by norepinephrine in vitro (*p<

0.01 vs. low dose; +p< 0.01 vs. baseline; ANOVA);

TRAP= Thrombin receptor activating protein inducing

maximal stimulation. (Doctoral thesis of Christoph

Tschuor)

Fig. 3

Dose- dependent decrease in production of radical

oxygen species (ROS) in isolated neutrophils from

healthy controls stimulated by norepinephrine in vitro

(*p< 0.01 vs. baseline; +p< 0.01 vs. norepinephrine;

ANOVA); PMA= Phorbol 12-myristate 13-acetate

(positive control). (Doctoral thesis of Martina Tanner)

Successful establishment of bed side BIS EEG in patients with severe traumatic

brain injury was the fi rst step in characterizing drug potencies of routinely

applied sedatives with the aim of using a sedation depth- targeted adaptation of

drug dosage, thereby possibly avoiding drug over- and underdosage. This will

be complemented by analysis of specifi c pharmacodynamic alterations in conjunction

with the analysis of drug concentrations and their active metabolites.


Achievements 2005

� Establishment of a laboratory for the Division of Surgical Intensive Care Medicine

� Approval by the local Ethics Committee

� Approval of a HPLC with an autosampler, a fluorescence and multiwave

detector to perform analysis of various mediators and markers of tissue

damage and determine drug concentrations

� Initiation of in house collaborations to determine changes in arterio- jugular

venous differences in oxycholesterol (a marker for cerebral cholesteral

degradation and membrane damage), amino acids, P-Selectin (a marker

for activated thrombocytes), and changes in functional activation of isolated

neutrophils and platelets

� John F. Stover was appointed executive member of the European Brain

Injury Consortium (EBIC)

Collaborations:

� PD Dr. med. Marius Keel, Division of Trauma Surgery, University Hospital Zuerich

� Dr. rer. nat. Riem Ha, Department of Internal Medicine, University Hospital Zuerich

� Dr. med. Lars Asmis, Institute for Clinical Hematology, University Hospital Zuerich

� PD Dr. Guido Wanner, Division of Trauma Surgery, University Hospital Zuerich

� PD Dr. Katharina Rentsch, Institute for Clinical Chemistry, University Hospital

Zuerich

� Dr. med. Oliver W. Sakowitz and Prof. Dr. Andreas W. Unterberg, Department

of Neurosurgery, University Hospital Heidelberg

� Dr. med. Ulrich W. Thomale, Department of Neurosurgery, Charite, Berlin

Selected references:

� Stover JF, Kempski OS. Anesthesia increases circulating glutamate in

neurosurgical patients. Acta Neurochir (Wien). 2005; 147 (8): 847- 853.

� Schaser KD, Puhl G, Vollmar B, Menger MD, Stover JF, Kohler K, Neuhaus

P, Settmacher U. In vivo imaging of human pancreatic microcirculation and

pancreatic tissue injury in clinical pancreas transplantation. Am J Transplant.

2005; 5 (2): 341-350.

� Schaser KD, Bail HJ, Schewior L, Stover JF, Melcher I, Haas NP, Mittlmeier

T. Acute effects of N-acetylcysteine on skeletal muscle microcirculation follo

wing closed soft tissue trauma in rats. J Orthop Res. 2005; 23 (1): 231- 241.

� Keel M, Mica L, Stover J, Stocker R, Trentz O, Harter L. Thiopental-induced

apoptosis in lymphocytes is independent of CD95 activation. Anesthesiology.

2005; 103 (3): 576- 584.

� Stover JF, Steiger P, Stocker R. Treating intracranial hypertension in patients

with severe traumatic brain injury during neurointensive care. Eur J Trauma

2005; 31 (4): 308- 330


70

Boris

Leskosek

Vlasta

Strohmeier

Astrid Morger

Alush Avdyli

3. Services

Services

3.1 Surgical skill laboratories

Histology

Photo and Graphic

Administration

Surgery requires a number of practical and manual skills that can be trained

in skill laboratories. In our facilities which are open to all members of the

department we provide a number of tools and machines in a surgical

enviroment. To perform operations under conditions similar to the clinical

situation, technical help is provided by our staff which is also responsible for

maintenance of our facilities.

3.2 Microsurgical laboratory

3.3 Histology

Surgical skill laboratories

Microsurgical laboratory

The microsurgery laboratory is a seperate section in which several operatingmicroscopes

are availabe to all members of the department requiring special

equipment. Maintenance of this laboratory includes all aspects of preparation

of surgical instruments, sterilization, and handling of waste materials. In

addition, an intravital microscope including video equipment is available. This

facility also provides for histological work-up.

The laboratory for Histology provides a histological work-up from preserved

specimen to sectioning and staining. The labortaory contains an embedding

machine, several microtomes and staining devices. Several techniques

including paraffin embedded, frozen and plastic embedded tissue can be

processed.


Nico Wick,

Photographer

Stefan Schwyter,

Scientific

Illustrator

Susanne Frehner,

Administration

Division of Surgical

Research

Lea Schütz-Cohen,

Photographer

Carol De Simio,

Scientific

Illustrator

3.4 Photo and graphic services

A quick, flexible, versatile and professional service.

We offer

� photographic documentation of patients

� technical photography, on location or in our well equipped studio

� reproductions from any original

� laserprint (up to A4) and inkjet (up to A3) on any material

� preparation of files for external printing

� layout of printing matters

� graphic and design of illustrations for papers and books

� construction and maintainance of websites

� maintainance of the digital image archives

3. 5 Administration

� Administrative office management

� Financial accounting of the Research Divison

� Organisation, planning and coordination of Workshops and vocational training

� Workshop, tutorials and seminars

� Quarterly reports preparation

� Meeting coordination for the head of the Research Division

� Personnel administration of the employees of the University Hospital

Zurich and the University Zurich


72

4. Events and Workshops at the Surgical

Research Division in 2005

Newly established Virtual Skills Lab

ZKF round table

Student sewing and injection classes

Boris Leskosek's celebration of 35 years service


5. Publications 2005

� Baaijens F, Bouten C, Hoerstrup S, Mol A, Driessen N, Boerboom R.

Functional tissue engineering of the aortic heart valve. Clin Hemorheol

Microcirc 2005;33:197-199.

� Egermann M, Goldhahn J, Schneider E. Animal models for fracture

treatment in osteoporosis. Osteoporos Int 2005;16 Suppl 2:S129-138.

� Giuliani M, Moritz W, Bodmer E, Dindo D, Kugelmeier P, Lehmann R,

Gassmann M, et al. Central necrosis in isolated hypoxic human pancreatic

islets: evidence for postisolation ischemia. Cell Transplant 2005;14:67-76.

� Graf R, Schiesser M, Reding T, Appenzeller P, Sun LK, Fortunato F,

Perren A, et al. Exocrine Meets Endocrine: Pancreatic Stone Protein and

Regenerating Protein-Two Sides of the Same Coin. J Surg Res 2005.

� Handschin AE, Trentz OA, Hoerstrup SP, Kock HJ, Wanner GA, Trentz

O. Effect of low molecular weight heparin (dalteparin) and fondaparinux

(Arixtra) on human osteoblasts in vitro. Br J Surg 2005;92:177-183.

� Keel M, Mica L, Stover J, Stocker R, Trentz O, Harter L. Thiopental-

induced apoptosis in lymphocytes is independent of CD95 activation.

Anesthesiology 2005;103:576-584.

� Kelm JM, Diaz Sanchez-Bustamante C, Ehler E, Hoerstrup SP, Djonov V,

Ittner L, Fussenegger M. VEGF profiling and angiogenesis in human

microtissues. J Biotechnol 2005;118:213-229.

� Kelm JM, Dionov V, Ittner LM, Hoerstrup SP, Fussenegger M. Design of

custom-shaped vascularized tissues using microtissue spheroids as

minimal building units. Tissue Engineering 2005.

� Mol A, Driessen NJ, Rutten MC, Hoerstrup SP, Bouten CV, Baaijens FP.

Tissue engineering of human heart valve leaflets: a novel bioreactor for

a strain-based conditioning approach. Ann Biomed Eng 2005;33:1778-

1788.

� Mol A, van Lieshout MI, Dam-de Veen CG, Neuenschwander S,

Hoerstrup SP, Baaijens FP, Bouten CV. Fibrin as a cell carrier in cardio

vascular tissue engineering applications. Biomaterials 2005;26:3113-

3121.

� Reding TV, Bimmler DR, Perren A, Sun LK, Fortunato F, Storni F, Graf

R. A selective COX-2 inhibitor suppresses chronic pancreatitis in an ani

mal model (WBN/Kob rats): significant reduction of macrophage infiltration

and fibrosis. Gut 2005.

� Schmidt D, Mol A, Neuenschwander S, Breymann C, Gossi M, Zund

G, Turina M, et al. Living patches engineered from human umbilical cord

derived fibroblasts and endothelial progenitor cells. Eur J Cardiothorac

Surg 2005;27:795-800.

� Selzner N, Selzner M, Jochum W, Amann-Vesti B, Graf R, Clavien PA.

Mouse livers with macrosteatosis are more susceptible to normothermic

ischemic injury than those with microsteatosis. J Hepatol 2006;44:694-

701.

� Sharma S, Zhu L, Yang SC, Zhang L, Lin J, Hillinger S, Gardner B, et al.

Cyclooxygenase 2 inhibition promotes IFN-gamma-dependent enhance

ment of antitumor responses. J Immunol 2005;175:813-819.

� Sodian R, Fu P, Lueders C, Szymanski D, Fritsche C, Gutberlet M,

Hoerstrup SP, et al. Tissue engineering of vascular conduits: fabrication

of custom-made scaffolds using rapid prototyping techniques. Thorac

Cardiovasc Surg 2005;53:144-149.


74

� Trentz OA, Handschin AE, Bestmann L, Hoerstrup SP, Trentz OL, Platz A.

Influence of brain injury on early posttraumatic bone metabolism. Crit

Care Med 2005;33:399-406.

� Zweifel M, Breu K, Matozan K, Renner E, Welle M, Schaffner T, Clavien

PA. Restoration of hepatic mast cells and expression of a different mast

cell protease phenotype in regenerating rat liver after 70%-hepatectomy.

Immunol Cell Biol 2005;83:587-595.


Cardiac Surgery

6. Grants 2005

Grants Title of Project Project Leader

Federal Comission for In vivo evaluation of growth in tissue engineered Prof. Zünd

Technology and Innovation (CTI/KTI) arteries

Prof. Hoerstrup

SNF (NF46) Cell and matrix evaluation in tissue engineering Prof. Hoerstrup

EU Grant Framework Program 6

(BIOSYS)

Regenerative medicine in cardiovascular surgery Prof. Hoerstrup

SYMETIS Research Grant Cardiovascular tissue engineering

Prof. Zünd

Prof. Hoerstrup

CO-ME Robotics in cardiovascular surgery

Prof. Zünd

PD Dr. Grünenfelder

BMBF Cell preservation methods for cardiovascular surgery

Prof. Hoerstrup

Visceral & Transplant Surgery

Grants Title of Project Project Leader

Hepatobiliary laboratory

SNF Soluble mediators and cellular receptors in the ischemic liver Prof. Clavien

SNF Small-for-size liver transplantation: platelets and platelet-derived

serotonin in the ischemic and regenerating liver

Prof. Clavien

Bonizzi-Theler Die Schutzmechanismen in der ischämisch-präkonditionierten

Leber

Dr. Rüdiger/ Prof. Clavien

Zürcher Krebsliga Pfortaderligatur Prof. Clavien

UBS Role of Non-Parenchymal Cells for the Induction of Regeneration PD Dr. Selzner

in the Steatotic Liver.

Norvartis Stiftung für Hypothermic oxygenated perfusion extracorporeal of the rat liver PD Dr. Ph. Dutkowski

Medizinisch-Biologische in non heart beating donors after cold storage

Forschung

Pancreatitis laboratory

SNF Pancreatic Thread Proteins - Key Factors of a Sealing System PD Dr. Graf

for Epithelial Lesions in Pancreatic Ducts

Waring Analysis of inflammatory suppression in a rat model (WBN/Kob) PD Dr. Graf

of chronic pancreatitis

Waring Investigation of acute and chronic pancreatitis PD Dr. Bimmler

Helen Biber Fonds Suppression of inflammation in an animal model of chronic PD Dr. R. Graf

pancreatitis (WBN/Kob rat)

Islet-Transplantation laboratory

SNF Ischemic preconditioning and gene therapeutic strategies to improve PD Dr. Weber/Dr. Moritz

islet cell engraftment in human pancreatic islet transplantation.

Hermann-Klaus Einfluss der Hypoxie auf den Apoptotischen Zelltod im Rahmen PD Dr. Weber/Dr. Moritz

der Inseltransplantation

Olga Mayenfisch Präkonditionierung PD Dr. Weber/Dr. Moritz

Hartmann Müller Die Verwendung eines natürlich vorkommenden anti-mikrobiellen PD Dr. Weber/Dr. Moritz

Peptids zur Verbesserung des Engraftments bei intrahepatischer

Inselzelltransplantation


76

Trauma Surgery

Grants Title of Project Project Leader

SNF Wound Healing in Vacuum Assisted Closure-Treated Patients Dr. Keel, Dr. Härter,

after Trauma: Implications of Neutrophil Activation for

Accelerated Angiogenesis

Dr. Labler

Heuberg Stiftung Modulation der Apoptose durch MAP-Kinasen in neutrophilen

Granulozyten: Bedeutung in der Pathogenese der systemischen

Entzündungsreaktion (SIRS) nach Trauma

Dr.Keel, Dr. Härter

AO Research Foundation Assessment of soft tissue and periosteal microcirculation in severely Dr. Wanner

open fractures using orthogonal polarozation spectral imaging

Stiftung für wissenschaftl. Nichterythroide Wirkungen von humanem rekombinaten Erythro- Dr. Wanner

Forschung der Universität poietin in der Traumatologie und rekonstruktiven Chirurgie der

Zürich Extremitäten

Novartis

AO Research Commission

Hartmann Müller-Stiftung

Sanofi Synthelabo

Modulation of Host Responses to Bacterial Endotoxin

PD Dr. Heinzelmann

Plastic Hand & Reconstructive Surgery

Grants Title of Project Project Leader

SUVA und Jubiläumsstiftung Tissue Engineering Dr. Wedler

Swiss Life Tissue Engineering Dr. Wedler


Thoracic Surgery

Grants Title of Project Project Leader

Krebsliga Pharmacokinetic Study after Intrapleural Topical Application

of Chemotherapeutic Agent for malignant Pleuramesothelioma

in Immune Competent Rat Model

PD Dr. Lardinois

Hartmann Müller-Stiftung Pharmacokinetic Study after Intrapleural Topical Application of

Chemotherapeutic Agent for malignant Pleuramesothelioma in

Immune Competent Rat Model

PD Dr. Lardinois

Hartmann-Müller-Stiftung Sildenafil verlängert das Graft Überleben in einem Grosstier-

Transplantations Modell

Dr. Cardell

Olga Mayenfisch - Melatonin (MLT) Melatonin in der Transplantationsimmunologie PD Dr. Korom

Krebsliga Zürich Adjuvante intrapleurale Spüllösung nach Pleuropneumonektomie Dr. Schmitt-Opitz

beim malignen Pleuramesotheliom

Krebsliga Epstein Barr virus-induced molecule 1 ligand chemokine in

lung cancer therapy

Dr. Hillinger

Sassella-Stiftung Epstein Barr virus-induced molecule 1 ligand chemokine in

lung cancer therapy

Dr. Hillinger

NSAID Study Assessment of the degree of pleurodesis after pleural mechanical PD Dr. Lardinois

abrasion and administration of COX-2 selective inhibitors and

nitric oxide - relaesing NSAI-drugs in comparison to classical

NSAIDs in a pig model

Hartmann Müller-Stiftung Immunsuppressive und zytostatische Wirkung von Gemzitabine Dr. Jung

in tumorinokkulierten Empfängern perfundierter Organtransplanatate

SNF The T cell costimulatory antigen CD26/Dipeptidyl Peptidase IV

(DPP IV) in acute rejection of lung allografts

PD Dr. Korom

SNF Experimental tracheal reconstruction using different developed Dr. Yang/

extent tissue-engineered cylinder construct Prof. Weder

EMDO-Stiftung Entwicklung eines in-vivo-Bioreaktors zur Reepithelialisierung

einer Tissue-engineerten Neo-Trachea

Dr. Hillinger


78

7. Awards 2005

� Pfizer Forschungs-Preis.Dave H. Minimal invasive Chirurgie für die Behebung

von angeborenen Herzfehler. Stiftung Pfizer Forschungs-Preis, Zürich.

� Research Posterpreis, Universitätskinderklinik Zürich, Kadner A, Dave H,

Dodge-Khatami A, Balmer C, Bürki C, Bauersfeld U, Prêtre R. “Right

axillary incision: a cosmetically superior approach to repair a wide range

of cardiac defects”.

� Preis für den besten Vortrag: EVAR to reduce morbidity and mortality in

treatment of ruptured abdominal aortic aneurysms: a seven year experience

with bifurcated stent-grafts. 37. Jahrestagung der Österreichischen

Gesellschaft für Gefässchirurgie, Krems, Österreich, 13.-15. Oktober 2005.

� International Society for Vascular Surgery Award 2005: Vacuum assisted

closure system with direct contact to native arteries and/or vascular grafts

to improve the outcome of perivascular infection. 32nd Annual Vascular and

Endovascular Issues, Techniques and Horizons (VEITH) Symposium, N.Y.,

USA, 17.-20. November 2005.

� Best Oral Presentation, Dörthe Schmidt, 4th Clinical Day of Research,

Center for Clinical Research, University Hospital Zürich

� Hartmann Mülller Stiftung, Tissue engineering using prenatal progenitors,

Schmidt, D (2005)

� Auszeichnung durch die Deutschen Gesellschaft f. Herz- Thorax – und

Gefässchirurgie für die Implementierung von CIRS (Critical Incidence

Reporting System)

� Ph. Dutkowski, Research award from the Swiss Society of Surgery

� H. Heinrich, Research award from the Swiss Society of Surgery

� D. Dindo, Research award from the Swiss Society of Surgery

� PD Dr. Tian received a Poster Prize at 4th Clinical Day of Research,

Center for Clinical Research, University Hospital Zürich


80

Sponsors:

AOTEC AG

More magazines by this user
Similar magazines